JP4378909B2

JP4378909B2 - Biological cell culture control method, culture apparatus control apparatus, and culture apparatus

Info

Publication number: JP4378909B2
Application number: JP2002042510A
Authority: JP
Inventors: 良一芳賀; 勝難波; 研天野
Original assignee: Hitachi Plant Technologies Ltd
Current assignee: Hitachi Plant Technologies Ltd
Priority date: 2002-02-20
Filing date: 2002-02-20
Publication date: 2009-12-09
Anticipated expiration: 2022-02-20
Also published as: JP2003235544A

Description

【０００１】
【発明の属する技術分野】
本発明は、生体細胞の培養制御方法及び培養装置の制御装置並びに培養装置に関する。
【０００２】
【従来の技術】
生体の細胞を培養する場合においては、培養環境、すなわち培養槽内を培養に最適な条件に維持することが求められる。このため、溶存酸素濃度，ｐＨ，温度，撹拌速度、等を至適条件に維持することが行われている。
【０００３】
溶存酸素濃度を制御する方法として、特公昭６０−１８３９０号公報（特許第１５５２５６３号）がある。これは溶存酸素濃度を測定するセンサーを培養槽に設置して、その指示値に合わせて培養槽の運転条件を制御する方法である。
【０００４】
また、ｐＨを制御する方法として、特開昭５８−８１７８１号公報がある。これは培養槽気相部の炭酸ガス濃度を変化させることにより培養液のｐＨを調節する方法である。
【０００５】
また、撹拌速度を制御する方法として、特開平５−３０９６２号公報がある。これは培養槽内に培養液の粘度を測定するセンサーを設け、その指示値と回転速度から剪断応力を算出し生体の細胞を破壊するに至らない剪断応力の範囲で撹拌速度を調節する方法である。
【０００６】
これらの従来の手法は、予め設定した目標値を満足するよう制御するものであり、培養状況に対応して制御するものではなかった。
【０００７】
培養状況の異常を判定する方法として、特開平５−２４０６７３号公報がある。これは培養槽に設けた検出手段の指示値の標準値に対する相対偏位量を算出し、ファジー推論演算によって培養の正常度・異常度を計算して判定する方法である。
【０００８】
このような従来の手法では、予め入力した正常な培養状態での標準値をもとに培養の異常の有無を判定するものであり、刻々と変化する培養状況に的確に対応して判断を行うものではない。
【０００９】
【発明が解決しようとする課題】
培養状況は刻々と変化し、培養初期の生体細胞の濃度が低い状況では良好に制御されていたものが、培養によって生体細胞の濃度が高まることによって目標とする培養環境を維持できない状況の発生もありうる。通常の培養装置では計測手段を多数設置することはまれであり、培養槽内に不適切な培養環境が存在していたとしてもこれを検出することができない。また、培養の進行に伴って制御目標値の変更が必須な培養もある。
【００１０】
本発明の目的は、生体細胞の培養に適切な培養環境が確実に維持できていることを検証しつつ培養を行い、必要に応じて目標値の変更を適切に実施することができる生体細胞の培養制御方法及び培養装置の制御装置並びに培養装置を提供することにある。
【００１１】
【課題を解決するための手段】
本発明は、培養装置により生体細胞を培養する生体細胞の培養制御で、前記培養装置の運転状態を計測した計測値が予め設定された目標値に一致するよう制御を行い、前記計測値と前記培養装置から採取した培養液試料を分析して得た分析値とに基づき該目標値を変更することを特徴とする。
【００１２】
【発明の実施の形態】
本発明の実施の形態を図面を用いて以下詳細に説明する。図１は、生体の細胞を培養する培養装置の制御方法を示すブロック図である。
【００１３】
本発明の実施の形態による制御方法は、培養装置により生体細胞を培養する生体細胞の培養制御で、前記培養装置の運転状態を計測した計測値が予め設定された目標値に一致するよう制御を行い、前記計測値と前記培養装置から採取した培養液試料を分析して得た分析値とに基づき該目標値を変更するものである。
【００１４】
個別制御手段２２は、予め設定された目標値と培養槽に設けた計測手段６より得られる計測値を比較して動作信号を操作手段２８に伝達して操作量を変更することにより、制御目標値にそれぞれの計測値が収束するよう自律的に制御動作を実行する第１のステップの制御を行う。
【００１５】
個別制御手段２２としては特に限定するものではなく、ｐＨ，溶存酸素濃度および温度等を制御量とするものが用いられる。なお、培養する生体の細胞が動物細胞である場合、制御量がｐＨである個別制御手段２２の操作手段は炭酸ガス供給弁およびポンプであり、それぞれ操作量は炭酸ガス供給量およびアルカリ注入量である。制御量が溶存酸素濃度である個別制御手段２２の操作手段は酸素供給弁および窒素供給弁であり、それぞれの操作因子は酸素供給量および窒素供給量である。制御量が温度である個別制御手段２２の操作手段は加温用ヒーター電流調節器または蒸気供給弁、および冷却水供給弁であり、操作因子はヒーターへの供給電力量または蒸気供給量、および冷却水供給量を操作量とする。個別制御手段２２については特に限定するものではなく、比例制御法，ＰＩＤ制御法等の公知のフィードバック制御手法を用いれば良い。なお、それぞれの制御目標値の設定・変更がコンピュータ２３によって行えることが好ましい。
【００１６】
コンピュータ２３は、計測手段６からの計測値と入力手段２７から入力される分析値とを用いて前記目標値の妥当性を検証し、記憶手段２４に格納された前回の検証過程と結果および過去の培養データと比較し、前記目標値の妥当性を検証して表示手段２５に表示するとともに、必要な場合には該目標値を変更し、および必要な場合には警報手段２６によって異常警報を出力する第２ステップの制御を行う。コンピュータ２３は下記▲１▼〜▲８▼の動作を実行する。
【００１７】
▲１▼計測手段６からの計測値と入力手段２７から入力される分析値とを用いて演算を行い、培養評価因子の算出と数時間〜数日後の予測値を算出する。
【００１８】
▲２▼計測手段６からの計測値と入力手段２７から入力される分析値とが現状の培養状況において妥当な数値であるかどうか、および▲１▼の演算結果が前回の予測値の許容範囲内にあるか否か判定する。許容範囲内にあるときは▲４▼に進む。許容範囲外であるときは▲３▼に進む。
【００１９】
▲３▼▲１▼の演算結果が記憶手段２４に保存されたデータベースの許容範囲内にあるか否か判定する。許容範囲内にあるときは▲４▼に進む。許容範囲外であるときは表示手段２５，警報手段２６により培養が異常状態にあることを表示し、▲５▼に進む。
【００２０】
▲４▼現状の培養状況とデータベースとを比較し、制御目標値の変更が必要かどうか判定する。すなわち、組み替えた遺伝子の発現制御が温度やｐＨ，溶存酸素，剪断応力によって行われる場合や発現誘導剤の添加で行われる場合に、現状の培養状況が遺伝子発現操作をすべき時期か否かを判定する。必要がないと判定した場合は▲５▼に進む。変更が必要と判定した場合は▲６▼に進む。
【００２１】
▲５▼現状の制御目標値で培養槽内での物質収支を演算し、培養槽内に不適切な環境の存在の有無を判定する。不適切な環境の存在があると判断された場合は▲６▼に進む。ないと判定された場合は▲１▼に戻り、第２のステップの制御を繰り返す。
【００２２】
▲６▼変更すべき制御因子とその制御目標値候補値を決定する。
【００２３】
▲７▼制御目標値候補値を用いて培養槽内での物質収支を演算し、培養槽内に不適切な環境形成の有無を判定する。不適切な環境が形成されると判断した場合は▲６▼に戻り、新たな制御目標値候補値を決定する。不適切な環境は形成されないと判定した場合は▲８▼に進む。
【００２４】
▲８▼個別制御手段の制御目標値の設定を変更する。▲１▼に戻り、終了の指令が出されるまで第２ステップの一連の制御動作を繰り返す。なお、安全性の確保の観点からは、目標値を変更するに際しては、予め登録された目標値変更の実施権限を付与された変更認定者の立会いを確認する動作と、該変更認定者の目標値変更認可を確認する動作を完了した後でなければ制御目標値の変更ができないようにすることが好ましい。
【００２５】
培養評価因子としては特に限定するものではないが、比増殖速度，生存率，基質消費速度，生産物生産速度，酸素消費速度，炭酸ガス生成速度、等を用い、必要に応じて他の因子を加える。培養の数時間〜数日後を予測する方法としては特に限定するものではなく、過去の培養データについてｐＨ，温度，酸素消費速度，基質濃度等の影響を多重解析法によって近似した実験式を用いて算出する方法等を用いれば良い。培養槽内での物質収支を演算する手法としては特に限定するものではなく、槽内の流れの乱流エネルギー散逸速度εを用いる乱流モデルを利用して流体力学的手法により解析する方法等を用いることができる。
【００２６】
以上のように、本発明の実施の形態による制御方法によれば、刻々と変化する培養状況に対応して目標値の変更を適切に実施することができ、かつ培養に適切な培養環境が確実に維持できていることを検証しつつ培養を行うことが可能となり、安全で確実な培養が可能となる。
【００２７】
図２は、本発明の実施の形態による制御装置の一実施例の動作を説明するフロー図である。制御対象の培養装置には溶存酸素濃度，ｐＨ，温度の計測手段を設置し、それぞれの計測値に基づいて予め定めた制御目標値に収束させるべく各手段毎に個別制御手段２２が設けられておりそれぞれ独立した制御操作を実施する。なお、図２に記載した計測手段とは、培養装置に設置した計測手段のいずれか一つを例として示すものであって、計測手段毎に図２のフローが独立的に実行される。また、計測手段としては前記の手段に限定されるものではなく、培養液濁度等の他の手段を加えても良い。
【００２８】
培養開始の信号が入力されることにより制御が開始される。制御フローの各ステップの動作を以下に説明する。
【００２９】
Ｓ１１：培養装置に設置した溶存酸素濃度，ｐＨ，温度の計測手段により、それぞれの計測値を得る。
【００３０】
Ｓ１２：個別制御手段において、それぞれの計測値が予め設定された制御目標値に一致するかどうか判定する。一致する場合はＳ１１に戻る。一致しない場合はＳ１３に進む。
【００３１】
Ｓ１３：Ｓ１２において、制御目標値に一致していないと判断された場合には、制御目標値に収束するよう、それぞれの個別制御手段において操作手段に対して動作信号を伝達し、操作量を変更する。それぞれの個別制御手段における制御手法としては特に限定するものではなく、ＯＮ／ＯＦＦ制御法，比例制御法，
ＰＩＤ制御法等の公知の手法を用いることができる。変更後、Ｓ１１に戻る。なお、各制御手段での操作量としては、下記のものが用いられる。
【００３２】
ｐＨ：通気ガス中の炭酸ガス供給量の増減、および酸性溶液またはアルカリ性溶液の注入量。
【００３３】
溶存酸素濃度：通気ガス中の酸素供給量の増減，培養液撹拌速度の増減，培養槽圧力の増減。
【００３４】
温度：ジャケット供給水温度の増減，冷却水供給速度の増減，加熱用電気ヒーター供給電力量の増減または加熱用蒸気供給量の増減。
【００３５】
動作Ｓ１１〜Ｓ１３は、終了命令が発せられるまで反復して実行される。反復の周期は培養する生体細胞の特性、および培養装置の動特性をもとに適宜決定されるが、概ね１秒〜１０分の範囲で実施される。
【００３６】
Ｓ２１：培養装置より生体細胞の培養液を無菌的に採取する。採取の手法は特に限定するものではなく、作業者が手作業で採取しても自動採取装置を用いても良い。
【００３７】
Ｓ２２：Ｓ２１で採取した培養液試料について必要な分析を行う。分析項目としては細胞濃度，細胞生存率，基質物質であるグルコースおよびグルタミンの濃度，代謝物質である乳酸，アンモニア，乳酸脱水素酵素および目的生産物の濃度、のいずれか１つ以上を実施するのが好ましいが、特にこれらに限定するものではない。
【００３８】
Ｓ２３：Ｓ２２で得た分析値をコンピュータに入力する。
【００３９】
Ｓ２４：Ｓ２１の動作を実施する時点で、Ｓ１１の計測データをコンピュータに取込む。
【００４０】
Ｓ２５：Ｓ２４で取込んだ計測データおよびＳ２３で入力した分析データをもとに演算を行い、培養評価因子の算出と数時間〜数日後の予測値を算出する。
【００４１】
Ｓ２６：Ｓ２５での解析結果をもとに、培養が正常に行われているか検証を実施する。すなわち、計測値と分析値とが現状の培養状況において妥当な数値であるかどうか、およびＳ２５の演算結果が前回の予測値の許容範囲内にあるか否か判定する。許容範囲内にあるときはＳ２９に進む。許容範囲外であるときはＳ27に進む。
【００４２】
Ｓ２７：Ｓ２５の演算結果が過去の培養データで構成されたデータベースでの許容範囲にあるかどうかを判定する。図３は総細胞濃度と生細胞濃度についてのＳ２５での演算結果と過去の培養データの関係を表す表示画面の一例を示す。上限値４３および下限値４４とで挟まれた領域が許容範囲である。本表示例では培養経過点４１、および予測値４２ともに許容範囲内にあることから、培養は良好に行われていると判断される。なお、許容範囲は正常に行われたと判断された過去の培養データをもとに算出する。本実施例ではデータベースの８０％が含まれる領域として決定した。許容範囲内にあるときはＳ２９に進む。許容範囲外であるときはＳ２８に進む。
【００４３】
Ｓ２８：現状の培養状況が異常であることを告知する異常警報を表示する。
Ｓ２９に進む。
【００４４】
Ｓ２９：現状の培養状況とデータベースとを比較し、制御目標値の変更が必要かどうか判定する。必要がないと判定した場合はＳ３０に進む。変更が必要と判定した場合はＳ３１に進む。
【００４５】
Ｓ３０：現状の制御目標値で培養槽内での物質収支を演算し、培養槽内に不適切な環境の存在の有無を判定する。不適切な環境の存在があると判断された場合はＳ３１に進む。ないと判定された場合はＳ２１に戻り、第２ステップの制御を繰り返す。
【００４６】
Ｓ３１：変更すべき制御因子とその制御目標値候補値を決定する。
【００４７】
Ｓ３２：制御目標値候補値を用いて培養槽内での物質収支を演算し、培養槽内に不適切な環境形成の有無を判定する。不適切な環境が形成されると判断した場合はＳ３１に戻り、新たな制御目標値候補値を決定する。不適切な環境は形成されないと判定した場合はＳ３５に進む。
【００４８】
Ｓ３５：個別制御手段の制御目標値を上記目標値候補値に変更する。Ｓ２１に戻り、終了の指令が出されるまで第２ステップの一連の制御動作を繰り返す。
【００４９】
本実施の形態の制御装置によれば、刻々と変化する培養状況に対応して目標値の変更を適切に実施することができ、かつ培養に適切な培養環境が確実に維持できていることを検証しつつ培養を行うことが可能となり、安全で確実な培養を実施できる。
【００５０】
図４は、本発明の実施の形態による制御装置の他の実施例の動作を説明するフロー図である。図５は、本発明の実施の形態による動物細胞を培養するための培養装置の１例を示す。
【００５１】
本培養装置は、培養槽１および制御装置２１とで構成される。また、図１中には図示していないが、培養設備には不可欠である、空気，酸素，窒素および炭酸ガス等のガス供給設備，温水冷水供給設備，蒸気供給設備および給排水設備を具備している。
【００５２】
また、計測手段６，７，８および９、並びに個別制御手段２２については、実装置では検出項目毎または制御項目毎に１つの検出手段が用いているが、図１中には簡略化の為それぞれ１つのみ記載した。
【００５３】
培養槽１は断面で表わしている。培養槽１内に張り込まれた培養液２は、駆動用モーター３により駆動される攪拌機４で撹拌され、均一に混合される。培養に必要な酸素は、酸素含有ガスを槽底部に配置されたスパージャー５から液中に供給する液中通気法と槽上部気相部に通気する上面通気法の二つの方法により供給される。
【００５４】
培養槽１には、計測手段６，供給ガスを計測する計測手段７および８，計測手段９を具備している。培養液２の性状を計測する計測手段６，排気ガスを計測する計測手段９および駆動用モーター３よりｐＨ，溶存酸素濃度，温度，撹拌速度，培養液濁度，排ガス中酸素濃度および排ガス中炭酸ガス濃度等の計測値１５を得る。計測値１５は後述するコンピュータ２３に伝送される。また、ｐＨ，溶存酸素濃度および温度の計測値については個別制御手段２２にも直接伝送される。計測手段７および８は、供給ガスおよび中和用アルカリ液の供給量を計測しかつ供給量調節する機能を有しており、総通気，空気，酸素，窒素，炭酸ガスの各供給量およびアルカリ供給量等の計測値１７を得る。各計測データ値は後述するコンピュータ２３に伝送される。
【００５５】
培養槽１には、培養液２の一部を採取する試料採取ライン１０を設けてあり、培養中に無菌的に培養液の一部を分析用の試料１１として採取することができる。試料１１は各種の分析装置１２に供され、細胞濃度，細胞生存率，グルコース濃度，乳酸濃度，アンモニア濃度，グルタミン濃度，乳酸脱水素酵素活性濃度および目的生産物濃度等の分析値１６を得る。分析値１６は入力手段２７を用いてコンピュータ２３に入力される。
【００５６】
制御装置２１は、図１に示す本発明の実施の形態による制御方法を実施するための機能を有するものであり、個別制御手段２２，コンピュータ２３，記憶手段２４，表示手段２５および警報手段２６とを具備する。
【００５７】
本制御装置２１は計測値１５を用いて予め設定された目標値を目標として培養制御を行う第一の制御手段と、計測値１５，計測値１７および分析値１６とを用いて前記目標値の妥当性を検証し、該検証過程と結果、前記データベースに格納された前回の検証過程と結果および過去の培養データと比較し、前記目標値の妥当性を検証して必要な場合には該目標値を変更し、および必要な場合には異常警報を出力する第二の制御手段を備えている。そしてこの第一及び第二の制御手段である２段階の制御を行う。
【００５８】
個別制御手段２２は前記第１ステップの制御を実行する第一の制御手段であり、ｐＨ，溶存酸素濃度および温度を制御量とする３基が設けられている。それぞれの個別制御手段は、それぞれ直接伝送されるｐＨ，溶存酸素濃度および温度の計測値が、あらかじめ設定された制御目標値にそれぞれの計測値が収束するよう操作手段２８に動作信号を伝達し、自律的に制御動作を実行する。なお、制御量がｐＨである個別制御手段２２の操作手段は炭酸ガス供給弁およびポンプであり、それぞれ操作量は炭酸ガス供給量およびアルカリ注入量である。
【００５９】
制御量が溶存酸素濃度である個別制御手段２２の操作手段は酸素供給弁および窒素供給弁であり、それぞれの操作因子は酸素供給量および窒素供給量である。制御量が温度である個別制御手段２２の操作手段は加温用ヒーター電流調節器または蒸気供給弁、および冷却水供給弁であり、操作因子はヒーターへの供給電力量または蒸気供給量、および冷却水供給量を操作量とする。個別制御手段２２については特に限定するものではなく、比例制御法，ＰＩＤ制御法等の公知のフィードバック制御手法を用いる。なお、それぞれの制御目標値の設定・変更がコンピュータ２３によって行える。
【００６０】
コンピュータ２３は前記第２ステップの制御を実行する第二の制御手段であり、下記▲１▼〜▲８▼の動作を実行する。
【００６１】
▲１▼計測値１５，計測値１７および分析値１６を用いて演算を行い、培養評価因子の算出と数時間〜数日後の予測値を算出する。演算結果をデータベースに格納する。
【００６２】
▲２▼計測値１５，計測値１７および分析値１６が現状の培養状況において妥当な数値であるかどうか、および▲１▼の演算結果が前回の予測値の許容範囲内にあるか否か判定する。許容範囲内にあるときは▲４▼に進む。許容範囲外であるときは▲３▼に進む。判定結果をデータベースに格納する。
【００６３】
▲３▼▲１▼の演算結果が記憶手段２４に保存されたデータベースの許容範囲内にあるか否か判定する。許容範囲内にあるときは▲４▼に進む。許容範囲外であるときは表示手段２５，警報手段２６により培養が異常状態にあることを表示し、▲５▼に進む。判定結果をデータベースに格納する。
【００６４】
▲４▼現状の培養状況とデータベースとを比較し、制御目標値の変更が必要かどうか判定する。必要がないと判定した場合は▲５▼に進む。変更が必要と判定した場合は▲６▼に進む。判定結果をデータベースに格納する。
【００６５】
▲５▼現状の制御目標値を用いて培養槽内での物質収支を演算し、溶存酸素濃度，ｐＨ，温度，剪断応力、等について不適切な環境が培養槽内に存在するかどうかを判定する。不適切な環境が存在すると判断された場合は▲６▼に進む。ないと判定された場合は▲１▼に戻り、第２ステップの制御を繰り返す。判定結果をデータベースに格納する。
【００６６】
▲６▼変更すべき制御因子とその制御目標値候補値を決定する。
【００６７】
▲７▼制御目標値候補値を用いて▲５▼と同様な培養槽内での物質収支を演算し、培養槽内に不適切な環境の有無を判定する。不適切な環境が形成されると判断した場合は▲６▼に戻り、新たな制御目標値候補値を決定する。不適切な環境はないと判定した場合は▲８▼に進む。判定結果をデータベースに格納する。
【００６８】
▲８▼個別制御手段の制御目標値の設定を変更する。▲１▼に戻り、終了の指令が出されるまで第２ステップの一連の制御動作を繰り返す。なお、目標値を変更するに際しては、予め登録された目標値変更の実施権限を付与された変更認定者の立会いを確認する動作と、該変更認定者の目標値変更認可を確認する動作の完了を必要とする。制御目標値の変更値，立会いの変更認定者名，変更時間の情報をデータベースに格納する。
【００６９】
第２ステップの制御は、上記▲１▼での演算に必要な分析値の入力によって開始される。第２ステップの制御の頻度は特に限定するものではないが、培養対象が動物細胞である場合には通常１時間〜２４時間の間隔で実行される。また、培養対象が微生物である場合には通常３０分〜２４時間の間隔で実行される。何れも、培養槽から採取された培養液の分析に要する時間を勘案して適宜決定される。
【００７０】
本実施の形態の培養装置によれば、刻々と変化する培養状況に対応して目標値の変更を適切に実施することができ、かつ培養に適切な培養環境が確実に維持できていることを検証しつつ培養を行うことが可能となり、安全で確実な培養を実施できる。
【００７１】
つまり、本発明の実施の形態によると、生体細胞の培養を行うことにより、刻々と変化する培養状況に対応して培養槽内の環境を培養に好適な条件に維持することが可能となり、安全で確実な培養を実施できる。また、培養に適切な培養環境が確実に維持できていることを検証しつつ培養を行うことから、培養によって生産される有用物質の安全性の検証が容易となる。さらに、培養における培養槽の計測値，培養液の分析値，検証の結果，目標値の変更時間，承認者等の情報を時系列的培養データベースとして記憶手段に格納しておくことにより、事後に行う製品の安全性にかかわる検証が容易となる。
【００７２】
【発明の効果】
本発明によると、生体細胞の培養に適切な培養環境が確実に維持できていることを検証しつつ培養を行い、必要に応じて目標値の変更を適切に実施することができるという効果を奏する。
【図面の簡単な説明】
【図１】本発明の制御方法の一実施例を示すブロック図である。
【図２】本発明の制御装置の一実施例の動作を示すフロー図である。
【図３】本発明の制御装置の一実施例における表示の一例を示す図である。
【図４】本発明の制御装置の他の実施例の動作を示すフロー図である。
【図５】動物細胞を培養対象とする本発明の培養装置の一実施例を示す概要図である。
【符号の説明】
１…培養槽、２…培養液、３…駆動用モーター、４…攪拌機、６，７，８，９…計測手段、２１…制御装置、２２…個別制御手段、２３…コンピュータ、２４…記憶手段、２５…表示手段、２６…警報手段、２７…入力手段。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a biological cell culture control method, a culture apparatus control apparatus, and a culture apparatus.
[0002]
[Prior art]
In the case of culturing living cells, it is required to maintain the culture environment, that is, the inside of the culture tank, under the optimum conditions for the culture. For this reason, it is performed to maintain the dissolved oxygen concentration, pH, temperature, stirring speed, and the like under optimum conditions.
[0003]
As a method for controlling the dissolved oxygen concentration, there is JP-B-60-18390 (Patent No. 1552563). This is a method in which a sensor for measuring the dissolved oxygen concentration is installed in a culture tank, and the operating conditions of the culture tank are controlled in accordance with the indicated value.
[0004]
JP-A-58-81781 discloses a method for controlling pH. This is a method of adjusting the pH of the culture solution by changing the carbon dioxide concentration in the gas phase part of the culture tank.
[0005]
Japanese Laid-Open Patent Publication No. 5-30962 discloses a method for controlling the stirring speed. This is a method in which a sensor for measuring the viscosity of the culture solution is installed in the culture tank, and the shear rate is calculated from the indicated value and the rotational speed, and the stirring speed is adjusted within the range of shear stress that does not lead to destruction of living cells. is there.
[0006]
These conventional methods are controlled so as to satisfy a preset target value, and are not controlled according to the culture state.
[0007]
Japanese Patent Laid-Open No. 5-240673 discloses a method for determining abnormalities in culture conditions. This is a method of calculating a relative deviation amount with respect to a standard value of an indication value of a detection means provided in a culture tank, and calculating and determining the normality / abnormality of culture by fuzzy inference calculation.
[0008]
In such a conventional method, the presence or absence of culture abnormality is determined based on a standard value in a normal culture state inputted in advance, and a determination is made in response to the constantly changing culture situation. It is not a thing.
[0009]
[Problems to be solved by the invention]
The culture situation changes from moment to moment, and it was well controlled when the concentration of living cells at the initial stage of culture was low, but there was also a situation where the target culture environment could not be maintained due to the increase in the concentration of living cells due to cultivation. It is possible. In a normal culture apparatus, it is rare to install a large number of measuring means, and even if an inappropriate culture environment exists in the culture tank, this cannot be detected. In addition, there is a culture in which the control target value must be changed as the culture progresses.
[0010]
An object of the present invention is to perform a culture while verifying that a culture environment suitable for the culture of biological cells can be reliably maintained, and to appropriately change the target value as necessary. An object of the present invention is to provide a culture control method, a control apparatus for the culture apparatus, and a culture apparatus.
[0011]
[Means for Solving the Problems]
The present invention performs control so that a measurement value obtained by measuring the operating state of the culture apparatus matches a preset target value in the culture cell culture control for culturing a living cell by the culture apparatus, and the measurement value and the The target value is changed based on the analysis value obtained by analyzing the culture solution sample collected from the culture apparatus.
[0012]
DETAILED DESCRIPTION OF THE INVENTION
Embodiments of the present invention will be described below in detail with reference to the drawings. FIG. 1 is a block diagram showing a control method of a culture apparatus for culturing living cells.
[0013]
In the control method according to the embodiment of the present invention, the control is performed so that the measured value obtained by measuring the operating state of the culture apparatus matches the preset target value in the culture cell culture control for culturing the living cell by the culture apparatus. The target value is changed based on the measured value and the analysis value obtained by analyzing the culture fluid sample collected from the culture apparatus.
[0014]
The individual control means 22 compares the target value set in advance with the measurement value obtained from the measurement means 6 provided in the culture tank, transmits an operation signal to the operation means 28, and changes the operation amount. Control of the first step of autonomously executing the control operation is performed so that each measurement value converges to the value.
[0015]
The individual control means 22 is not particularly limited, and an individual control means having pH, dissolved oxygen concentration, temperature and the like as control amounts is used. When the living cells to be cultured are animal cells, the operation means of the individual control means 22 whose control amount is pH is a carbon dioxide supply valve and a pump, and the operation amounts are the carbon dioxide supply amount and the alkali injection amount, respectively. is there. The operation means of the individual control means 22 whose control amount is the dissolved oxygen concentration is an oxygen supply valve and a nitrogen supply valve, and the respective operation factors are the oxygen supply amount and the nitrogen supply amount. The operation means of the individual control means 22 in which the control amount is the temperature is a heater current regulator for heating or a steam supply valve, and a cooling water supply valve, and the operating factors are the amount of electric power or steam supply to the heater, and the cooling The amount of water supply is the operation amount. The individual control means 22 is not particularly limited, and a known feedback control method such as a proportional control method or a PID control method may be used. In addition, it is preferable that each control target value can be set and changed by the computer 23.
[0016]
The computer 23 verifies the validity of the target value using the measured value from the measuring unit 6 and the analysis value input from the input unit 27, and the previous verification process and result stored in the storage unit 24 and the past In addition to verifying the validity of the target value, the target value is verified and displayed on the display means 25. If necessary, the target value is changed. The second step of output is controlled. The computer 23 executes the following operations (1) to (8).
[0017]
(1) An operation is performed using the measured value from the measuring means 6 and the analysis value input from the input means 27, and the calculation of the culture evaluation factor and the predicted value after several hours to several days are calculated.
[0018]
(2) Whether the measured value from the measuring means 6 and the analytical value input from the input means 27 are appropriate values in the current culture state, and the calculation result of (1) is the allowable range of the previous predicted value It is judged whether it is in. If it is within the allowable range, proceed to (4). If it is outside the allowable range, proceed to (3).
[0019]
(3) It is determined whether the calculation result of (1) is within the allowable range of the database stored in the storage means 24. If it is within the allowable range, proceed to (4). When it is out of the allowable range, the display means 25 and alarm means 26 indicate that the culture is in an abnormal state, and proceed to (5).
[0020]
(4) The current culture state is compared with the database to determine whether it is necessary to change the control target value. In other words, when the expression control of the recombined gene is performed by temperature, pH, dissolved oxygen, shear stress, or by the addition of an expression inducer, whether or not the current culture status is the time when gene expression manipulation should be performed. judge. If it is determined that it is not necessary, proceed to (5). If it is determined that a change is necessary, proceed to (6).
[0021]
(5) The material balance in the culture tank is calculated with the current control target value, and the presence or absence of an inappropriate environment in the culture tank is determined. If it is determined that there is an inappropriate environment, proceed to (6). If it is determined that there is no, return to {circle around (1)} and repeat the control of the second step.
[0022]
(6) The control factor to be changed and its control target value candidate value are determined.
[0023]
(7) The material balance in the culture tank is calculated using the control target value candidate value, and the presence or absence of inappropriate environment formation in the culture tank is determined. If it is determined that an inappropriate environment is formed, the process returns to (6) and a new control target value candidate value is determined. If it is determined that an inappropriate environment is not formed, proceed to (8).
[0024]
(8) Change the setting of the control target value of the individual control means. Returning to {circle around (1)}, the series of control operations in the second step is repeated until an end command is issued. From the viewpoint of ensuring safety, when changing the target value, the operation of confirming the presence of a change authorized person who has been granted the authority to perform the target value change registered in advance, and the target of the change authorized person It is preferable that the control target value can be changed only after the operation for confirming the value change authorization is completed.
[0025]
Although the culture evaluation factor is not particularly limited, the specific growth rate, survival rate, substrate consumption rate, product production rate, oxygen consumption rate, carbon dioxide production rate, etc. are used. Add. The method for predicting several hours to several days after the culture is not particularly limited, and an experimental formula that approximates the influence of pH, temperature, oxygen consumption rate, substrate concentration, etc. on past culture data by a multiple analysis method is used. A calculation method or the like may be used. The method for calculating the mass balance in the culture tank is not particularly limited, and a method of analyzing by a hydrodynamic method using a turbulent flow model using a turbulent energy dissipation rate ε of the flow in the tank is used. Can be used.
[0026]
As described above, according to the control method according to the embodiment of the present invention, it is possible to appropriately change the target value in response to the constantly changing culture state and to ensure a culture environment suitable for culture. Therefore, it is possible to carry out the culture while verifying that it can be maintained in a safe, reliable culture.
[0027]
FIG. 2 is a flowchart for explaining the operation of an example of the control device according to the embodiment of the present invention. The culture apparatus to be controlled is provided with measuring means for dissolved oxygen concentration, pH, and temperature, and individual control means 22 is provided for each means so as to converge to a predetermined control target value based on each measured value. Each control operation is carried out independently. Note that the measurement means described in FIG. 2 shows any one of the measurement means installed in the culture apparatus as an example, and the flow of FIG. 2 is executed independently for each measurement means. The measuring means is not limited to the above means, and other means such as culture turbidity may be added.
[0028]
Control is started by inputting a culture start signal. The operation of each step of the control flow will be described below.
[0029]
S11: Each measured value is obtained by means of measuring the dissolved oxygen concentration, pH, and temperature installed in the culture apparatus.
[0030]
S12: In the individual control means, it is determined whether or not each measured value matches a preset control target value. If they match, the process returns to S11. If they do not match, the process proceeds to S13.
[0031]
S13: When it is determined in S12 that the control target value does not match, the operation signal is transmitted to the operation means in each individual control means so as to converge to the control target value, and the operation amount is changed. To do. The control method for each individual control means is not particularly limited, and is an ON / OFF control method, a proportional control method,
A known method such as a PID control method can be used. After the change, the process returns to S11. In addition, the following are used as the operation amount in each control means.
[0032]
pH: Increase / decrease in the supply amount of carbon dioxide gas in the aeration gas, and the injection amount of acidic solution or alkaline solution.
[0033]
Dissolved oxygen concentration: Increase / decrease in oxygen supply in the aeration gas, increase / decrease in culture solution agitation rate, increase / decrease in culture tank pressure.
[0034]
Temperature: Increase / decrease in jacket supply water temperature, increase / decrease in cooling water supply speed, increase / decrease in heating electric heater supply power amount, or increase / decrease in heating steam supply amount.
[0035]
Operations S11 to S13 are repeatedly executed until an end command is issued. The cycle of repetition is appropriately determined based on the characteristics of the living cells to be cultured and the dynamic characteristics of the culture apparatus, and is generally performed in the range of 1 second to 10 minutes.
[0036]
S21: A culture solution of living cells is aseptically collected from the culture apparatus. The collection method is not particularly limited, and an automatic collection device may be used even if an operator manually collects the sample.
[0037]
S22: Necessary analysis is performed on the culture solution sample collected in S21. Analysis items include cell concentration, cell viability, concentrations of substrate substances glucose and glutamine, and metabolites lactic acid, ammonia, lactate dehydrogenase and target product concentrations. However, it is not particularly limited to these.
[0038]
S23: The analysis value obtained in S22 is input to the computer.
[0039]
S24: When the operation of S21 is performed, the measurement data of S11 is taken into the computer.
[0040]
S25: An operation is performed based on the measurement data captured in S24 and the analysis data input in S23, and a culture evaluation factor is calculated and a predicted value after several hours to several days is calculated.
[0041]
S26: Based on the analysis result in S25, it is verified whether the culture is normally performed. That is, it is determined whether the measured value and the analyzed value are appropriate numerical values in the current culture state, and whether the calculation result of S25 is within the allowable range of the previous predicted value. When it is within the allowable range, the process proceeds to S29. If it is outside the allowable range, the process proceeds to S27.
[0042]
S27: It is determined whether or not the calculation result of S25 is within an allowable range in a database composed of past culture data. FIG. 3 shows an example of a display screen showing the relationship between the calculation result in S25 for the total cell concentration and the living cell concentration and the past culture data. An area between the upper limit value 43 and the lower limit value 44 is an allowable range. In this display example, since the culture progress point 41 and the predicted value 42 are within the allowable range, it is determined that the culture is being performed well. The allowable range is calculated based on past culture data determined to have been performed normally. In this embodiment, the area is determined to include 80% of the database. When it is within the allowable range, the process proceeds to S29. If it is outside the allowable range, the process proceeds to S28.
[0043]
S28: An abnormal alarm for notifying that the current culture state is abnormal is displayed.
Proceed to S29.
[0044]
S29: The current culture state is compared with the database, and it is determined whether it is necessary to change the control target value. If it is determined that it is not necessary, the process proceeds to S30. If it is determined that a change is necessary, the process proceeds to S31.
[0045]
S30: The mass balance in the culture tank is calculated with the current control target value, and the presence or absence of an inappropriate environment is determined in the culture tank. If it is determined that there is an inappropriate environment, the process proceeds to S31. If it is determined that there is not, the process returns to S21 and the control of the second step is repeated.
[0046]
S31: The control factor to be changed and the control target value candidate value are determined.
[0047]
S32: The material balance in the culture tank is calculated using the control target value candidate value, and the presence or absence of inappropriate environment formation in the culture tank is determined. If it is determined that an inappropriate environment is formed, the process returns to S31, and a new control target value candidate value is determined. If it is determined that an inappropriate environment is not formed, the process proceeds to S35.
[0048]
S35: The control target value of the individual control means is changed to the target value candidate value. Returning to S21, the series of control operations in the second step is repeated until an end command is issued.
[0049]
According to the control device of the present embodiment, it is possible to appropriately change the target value corresponding to the culturing situation that changes every moment, and that the culturing environment suitable for culturing can be reliably maintained. Culture can be performed while verifying, and safe and reliable culture can be performed.
[0050]
FIG. 4 is a flowchart for explaining the operation of another example of the control device according to the embodiment of the present invention. FIG. 5 shows an example of a culture apparatus for culturing animal cells according to an embodiment of the present invention.
[0051]
The main culture apparatus includes a culture tank 1 and a control device 21. Although not shown in FIG. 1, it is equipped with gas supply equipment such as air, oxygen, nitrogen and carbon dioxide, hot / cold water supply equipment, steam supply equipment and water supply / drainage equipment, which are indispensable for culture equipment. Yes.
[0052]
As for the measuring means 6, 7, 8 and 9, and the individual control means 22, one detection means is used for each detection item or for each control item in the actual apparatus, but for simplification in FIG. Only one of each was listed.
[0053]
The culture tank 1 is represented by a cross section. The culture solution 2 stuck in the culture tank 1 is stirred by a stirrer 4 driven by a driving motor 3 and mixed uniformly. Oxygen required for the culture is supplied by two methods: a submerged aeration method in which oxygen-containing gas is supplied into the solution from a sparger 5 disposed at the bottom of the tank, and a top aeration method in which the gas is supplied to the upper gas phase of the tank. .
[0054]
The culture tank 1 includes a measuring means 6, measuring means 7 and 8 for measuring the supply gas, and a measuring means 9. PH, dissolved oxygen concentration, temperature, stirring speed, culture solution turbidity, oxygen concentration in exhaust gas and carbon dioxide in exhaust gas from measuring means 6 for measuring the properties of the culture medium 6, measuring means 9 for measuring exhaust gas, and driving motor 3 A measured value 15 such as gas concentration is obtained. The measured value 15 is transmitted to a computer 23 described later. The measured values of pH, dissolved oxygen concentration, and temperature are also directly transmitted to the individual control means 22. The measuring means 7 and 8 have the function of measuring the supply amount of the supply gas and the neutralizing alkali solution and adjusting the supply amount. The supply amounts of total aeration, air, oxygen, nitrogen, carbon dioxide and alkali A measurement value 17 such as a supply amount is obtained. Each measurement data value is transmitted to a computer 23 described later.
[0055]
The culture tank 1 is provided with a sample collection line 10 for collecting a part of the culture solution 2, and a part of the culture solution can be collected aseptically as a sample 11 for analysis during culture. The sample 11 is supplied to various analyzers 12 to obtain analytical values 16 such as cell concentration, cell viability, glucose concentration, lactic acid concentration, ammonia concentration, glutamine concentration, lactate dehydrogenase activity concentration and target product concentration. The analysis value 16 is input to the computer 23 using the input means 27.
[0056]
The control device 21 has a function for carrying out the control method according to the embodiment of the present invention shown in FIG. 1, and includes an individual control means 22, a computer 23, a storage means 24, a display means 25, and an alarm means 26. It comprises.
[0057]
The control device 21 uses the first control means for controlling the culture with the target value set in advance using the measurement value 15 as a target, and the target value using the measurement value 15, the measurement value 17, and the analysis value 16. The validity is verified, the verification process and result, the previous verification process and result stored in the database and the previous culture data are compared, the validity of the target value is verified, and if necessary, the target Second control means is provided for changing the value and outputting an abnormal alarm if necessary. Then, two-stage control is performed as the first and second control means.
[0058]
The individual control means 22 is the first control means for executing the control of the first step, and three units having the control amounts of pH, dissolved oxygen concentration and temperature are provided. Each individual control means transmits an operation signal to the operation means 28 so that the measured values of pH, dissolved oxygen concentration, and temperature that are directly transmitted are converged to preset control target values, respectively. Perform control operations autonomously. The operation means of the individual control means 22 whose control amount is pH is a carbon dioxide supply valve and a pump, and the operation amounts are a carbon dioxide supply amount and an alkali injection amount, respectively.
[0059]
The operation means of the individual control means 22 whose control amount is the dissolved oxygen concentration is an oxygen supply valve and a nitrogen supply valve, and the respective operation factors are the oxygen supply amount and the nitrogen supply amount. The operation means of the individual control means 22 in which the control amount is the temperature is a heater current regulator for heating or a steam supply valve, and a cooling water supply valve, and the operating factors are the amount of electric power or steam supply to the heater, and the cooling The amount of water supply is the operation amount. The individual control means 22 is not particularly limited, and a known feedback control method such as a proportional control method or a PID control method is used. Each control target value can be set and changed by the computer 23.
[0060]
The computer 23 is a second control means for executing the control of the second step, and executes the following operations (1) to (8).
[0061]
(1) Calculation is performed using the measured value 15, the measured value 17 and the analyzed value 16 to calculate a culture evaluation factor and a predicted value after several hours to several days. Store the calculation result in the database.
[0062]
(2) Judgment whether the measured value 15, measured value 17 and analytical value 16 are appropriate values in the current culture state, and whether the calculation result of (1) is within the allowable range of the previous predicted value To do. If it is within the allowable range, proceed to (4). If it is outside the allowable range, proceed to (3). Store the judgment result in the database.
[0063]
(3) It is determined whether the calculation result of (1) is within the allowable range of the database stored in the storage means 24. If it is within the allowable range, proceed to (4). When it is out of the allowable range, the display means 25 and alarm means 26 indicate that the culture is in an abnormal state, and proceed to (5). Store the judgment result in the database.
[0064]
(4) The current culture state is compared with the database to determine whether it is necessary to change the control target value. If it is determined that it is not necessary, proceed to (5). If it is determined that a change is necessary, proceed to (6). Store the judgment result in the database.
[0065]
(5) Calculate the material balance in the culture tank using the current control target value, and determine whether there is an inappropriate environment in the culture tank for dissolved oxygen concentration, pH, temperature, shear stress, etc. To do. If it is determined that an inappropriate environment exists, proceed to (6). If it is determined that there is no, return to {circle around (1)} and repeat the control of the second step. Store the judgment result in the database.
[0066]
(6) The control factor to be changed and its control target value candidate value are determined.
[0067]
(7) Using the control target value candidate value, the material balance in the culture tank similar to (5) is calculated, and the presence or absence of an inappropriate environment in the culture tank is determined. If it is determined that an inappropriate environment is formed, the process returns to (6) and a new control target value candidate value is determined. If it is determined that there is no inappropriate environment, proceed to (8). Store the judgment result in the database.
[0068]
(8) Change the setting of the control target value of the individual control means. Returning to {circle around (1)}, the series of control operations in the second step is repeated until an end command is issued. When changing the target value, the operation for confirming the presence of the change authorized person who has been granted the authority to perform the target value change registered in advance and the operation for confirming the target value change approval of the change authorized person are completed. Need. Stores the information on the change value of the control target value, the witness change authorized person name, and the change time in the database.
[0069]
The control of the second step is started by inputting the analysis value necessary for the calculation in the above (1). The frequency of the control in the second step is not particularly limited, but when the culture target is an animal cell, it is usually executed at an interval of 1 hour to 24 hours. Moreover, when the culture target is a microorganism, it is usually performed at intervals of 30 minutes to 24 hours. In any case, the time is appropriately determined in consideration of the time required for analyzing the culture solution collected from the culture tank.
[0070]
According to the culture device of the present embodiment, it is possible to appropriately change the target value in response to the constantly changing culture situation, and to ensure that a culture environment suitable for culture can be reliably maintained. Culture can be performed while verifying, and safe and reliable culture can be performed.
[0071]
In other words, according to the embodiment of the present invention, it is possible to maintain the environment in the culture tank under conditions suitable for culture in response to the constantly changing culture conditions by culturing living cells. With this, reliable culture can be performed. In addition, since the culture is performed while verifying that the culture environment suitable for the culture can be reliably maintained, the safety of useful substances produced by the culture can be easily verified. Furthermore, by storing information such as culture tank measurement values, culture solution analysis values, verification results, target value change times, and approver information in the storage means as a time-series culture database, post-event Verification related to the safety of the product to be performed becomes easy.
[0072]
【The invention's effect】
Advantageous Effects of Invention According to the present invention, it is possible to perform culture while verifying that a culture environment suitable for culture of living cells can be reliably maintained, and it is possible to appropriately change target values as necessary. .
[Brief description of the drawings]
FIG. 1 is a block diagram showing an embodiment of a control method of the present invention.
FIG. 2 is a flowchart showing the operation of an embodiment of the control device of the present invention.
FIG. 3 is a diagram showing an example of display in an embodiment of the control device of the present invention.
FIG. 4 is a flowchart showing the operation of another embodiment of the control apparatus of the present invention.
FIG. 5 is a schematic view showing an embodiment of the culture apparatus of the present invention for culturing animal cells.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Culture tank, 2 ... Culture solution, 3 ... Motor for drive, 4 ... Stirrer, 6, 7, 8, 9 ... Measuring means, 21 ... Control apparatus, 22 ... Individual control means, 23 ... Computer, 24 ... Memory | storage means 25 ... display means, 26 ... alarm means, 27 ... input means.

Claims

A culture control method for culturing animal cells using a culture device, wherein the measurement value obtained by controlling the measurement value obtained by measuring the operating state of the culture device to match a preset target value, and the measurement value Both the value of the culture evaluation factor calculated using the analysis value obtained by analyzing the culture solution sample collected from the culture apparatus and the predicted value of the culture evaluation factor after several hours to several days are 1 to 24 hours. It is determined whether or not it is within the allowable range of the predicted value of the culture evaluation factor after several hours to several days after the previous prediction, and when it is out of the allowable range, the calculated value of the culture evaluation factor, Whether the predicted value of the culture evaluation factor after several hours to several days is within the allowable range of the numerical value of the culture evaluation factor calculated based on the culture data determined that the past culture was performed normally determine whether the allowable range of numerical values of the culture evaluation factors When it is determined to be out, dissolved oxygen concentration and calculates the mass balance in the culture tank by hydrodynamic analysis in the control target value of current, pH, temperature, the presence of inappropriate culture environment Shear Stress If it is determined that there is an inappropriate culture environment, calculate the mass balance in the culture tank by hydrodynamic analysis using multiple control target value candidate values. A culture control method for culturing animal cells , comprising: a second step of changing the target value to a control target value candidate value that repeatedly determines the presence or absence of an appropriate culture environment and does not form an inappropriate culture environment.

In the culture control method of a culture apparatus for culturing animal cells, a first step of performing control so that a measurement value from an operating state measurement means provided in the culture apparatus matches a preset target value; and the measurement value And the value of the culture evaluation factor calculated using the analysis value obtained by analyzing the culture fluid sample collected from the culture apparatus and the predicted value of the culture evaluation factor after several hours to several days are 1 hour to It is determined whether or not it is within the allowable range of the predicted value of the culture evaluation factor of the previous several hours to several days after predicted 24 hours ago. If it is out of the allowable range, the calculated value of the culture evaluation factor Whether the predicted value of the culture evaluation factor after several hours to several days is within the allowable range of the value of the culture evaluation factor calculated based on the culture data determined that the past culture was normally performed. or it determines the allowable range of numerical values of the culture evaluation factors When it is determined to be out of the inner it is dissolved oxygen concentration by calculating the mass balance in the culture tank by hydrodynamic analysis in the control target value of current, pH, temperature, inappropriate culture environment Shear Stress If there is an inappropriate culture environment, calculate the mass balance in the culture tank by hydrodynamic analysis using multiple candidate control target values. It is characterized by repeatedly determining the presence or absence of an inappropriate culture environment, and performing control in two stages of the second step of changing the target value to a control target value candidate value that does not form an inappropriate culture environment A culture control method for a culture apparatus for culturing animal cells.

3. The culture control method for a culture apparatus for culturing animal cells according to claim 2, wherein the measurement means provided in the culture apparatus includes culture turbidity, pH, dissolved oxygen concentration, temperature, stirring speed, aeration rate, air supply. A sample of culture fluid collected from the culture device that is at least one of a quantity, oxygen gas supply quantity, carbon dioxide supply quantity, nitrogen gas supply quantity, oxygen concentration in the injection gas, oxygen concentration in the exhaust gas, and alkali supply quantity Culturing animal cells characterized in that the analysis value obtained by analyzing is one or more of cell concentration, cell viability, glucose concentration, lactate concentration, ammonia concentration, glutamine concentration, and target product concentration Culture control method for a culture apparatus .

A control device for a culture device for culturing animal cells using a culture device, wherein the first control means performs control so that a measured value obtained by measuring the operating state of the culture device matches a preset target value; Both the value of the culture evaluation factor calculated using the measured value and the analysis value obtained by analyzing the culture solution sample collected from the culture apparatus and the predicted value of the culture evaluation factor after several hours to several days are 1 It is determined whether or not it is within the allowable range of the predicted value of the culture evaluation factor of the last several hours to several days after the time to 24 hours before, and if it is out of the allowable range, the calculated culture evaluation factor Both the value and the predicted value of the culture evaluation factor after several hours to several days are within the allowable range of the numerical value of the culture evaluation factor calculated based on the culture data determined that the past culture was performed normally. it is judged whether or not the allowable range of numerical values of the culture evaluation factors When it is determined to be out of the inner it is dissolved oxygen concentration by calculating the mass balance in the culture tank by hydrodynamic analysis in the control target value of current, pH, temperature, inappropriate culture environment Shear Stress If there is an inappropriate culture environment, calculate the mass balance in the culture tank by hydrodynamic analysis using multiple candidate control target values. Culture for cultivating animal cells having second control means for changing the target value to a control target value candidate value that repeatedly determines the presence or absence of an inappropriate culture environment and does not form an inappropriate culture environment Control device for the device.

In the control apparatus for a culture apparatus for culturing animal cells, a means for taking a measurement value from a measurement means provided in the culture apparatus, and a control for performing control so as to match a preset target value using the measurement value Means, a means for inputting an analysis value obtained by analyzing a culture solution sample collected from the culture apparatus, a computer for verifying the validity of the target value, culture data and verification of the animal cell A storage means for storing the process and results as a database, a display means for displaying the verification process and results, and a target set in advance using measurement values from the measurement means provided in the culture apparatus Both the first control means for controlling the culture with the value as a target, the value of the culture evaluation factor calculated using the measured value and the analysis value, and the predicted value of the culture evaluation factor after several hours to several days 1 hour to 24 It is determined whether it is within the allowable range of the predicted value of the culture evaluation factor after several hours to several days after the previous prediction, and when it is outside the allowable range, the calculated value of the culture evaluation factor, Whether the predicted value of the culture evaluation factor after several hours to several days is within the allowable range of the numerical value of the culture evaluation factor calculated based on the culture data determined that the past culture was performed normally determined, acceptable when it is judged to be out of the range, the dissolved oxygen concentration by calculating the mass balance in the culture tank by hydrodynamic analysis in the control target value of the current value of the culture evaluation factors , PH, temperature, and shear stress are determined whether or not an inappropriate culture environment exists, and if it is determined that an inappropriate culture environment exists, a plurality of control target value candidate values are used to determine the hydrodynamics. Inappropriate calculation of the material balance in the culture tank by analysis Determining repeatedly the presence or absence of a nutrient environment, for no improper culture environment is formed a control target value candidate value, to verify the validity of the target value, the verification process and results, previous stored in the database The second control that compares the verification process with the results and past culture data, verifies the validity of the target value, changes the target value if necessary, and outputs an abnormal alarm if necessary A control device for a culture apparatus for culturing animal cells , characterized in that the control is performed by the first and second control means.

6. The control apparatus for a culture apparatus for culturing animal cells according to claim 5, wherein the display means is configured to display past culture data, current culture data, and future expected culture progress together, And a control device for a culture device for culturing animal cells, characterized in that a warning is given when the expected culture progress deviates from past culture data.

In a culture apparatus for culturing animal cells, a means for taking in a measurement value from a measurement means provided in the culture apparatus, a control means for performing control so as to match a preset target value using the measurement value, Means for inputting an analysis value obtained by analyzing a culture fluid sample collected from the culture apparatus, a computer for verifying the validity of the target value, culture data and verification process and result of the animal cell Storage means for storing the data as a database, display means for displaying the verification process and results, and a target value set in advance using the measurement value from the measurement means provided in the culture apparatus As a first control means for performing culture control, the value of the culture evaluation factor calculated using the measured value and the analysis value, and the predicted value of the culture evaluation factor after several hours to several days are 1 hour. ~ 24 hours in advance It is determined whether it is within the allowable range of the predicted value of the culture evaluation factor after the previous several hours to several days, and when it is out of the allowable range, the calculated value of the culture evaluation factor and several hours to several It is determined whether both of the predicted values of the culture evaluation factor after the day are within the allowable range of the numerical value of the culture evaluation factor calculated based on the culture data determined that the past culture was normally performed, If it is determined that the value of the culture evaluation factor is outside the allowable range , the material balance in the culture tank is calculated using the current control target value, and the dissolved oxygen concentration, pH, temperature, and shear stress are not considered. The presence or absence of an appropriate culture environment is determined, and if it is determined that there is an inappropriate culture environment, the material balance in the culture tank is determined by hydrodynamic analysis using multiple candidate control target values. the calculates repeatedly the presence or absence of inappropriate culture environment For the control target value candidate value that does not form an inappropriate culture environment, the validity of the target value is verified, the verification process and result, the previous verification process and result stored in the database, and the past culture A second control means for comparing the data, verifying the validity of the target value, changing the target value if necessary, and outputting an abnormal alarm if necessary; And a culture apparatus for culturing animal cells, characterized by comprising a control device that performs control in two stages of the second control means.

8. The culture apparatus for culturing animal cells according to claim 7, wherein the display means is configured to display past culture data, current culture data, and future expected culture process together, and present and future predictions. A culture apparatus for cultivating animal cells, characterized in that a warning is given by an emphasis means that warns when the progress of culture that is performed deviates from past culture data.