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ES2558626T3 - Placenta post-parto de mamíferos, su uso y células madre placentarias de la misma - Google Patents

Placenta post-parto de mamíferos, su uso y células madre placentarias de la misma Download PDF

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Publication number
ES2558626T3
ES2558626T3 ES10182243.5T ES10182243T ES2558626T3 ES 2558626 T3 ES2558626 T3 ES 2558626T3 ES 10182243 T ES10182243 T ES 10182243T ES 2558626 T3 ES2558626 T3 ES 2558626T3
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stem cells
placental stem
cells
placenta
abc
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Robert J. Hariri
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Clarity Acquisition II LLC
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Anthrogenesis Corp
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48—Reproductive organs
    • A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3695—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the function or physical properties of the final product, where no specific conditions are defined to achieve this
    • A—HUMAN NECESSITIES
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    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
    • A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/02—Blood transfusion apparatus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A61P7/06—Antianaemics
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    • C12N5/0602—Vertebrate cells
    • C12N5/0603—Embryonic cells ; Embryoid bodies
    • C12N5/0605—Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
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    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
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    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0676—Pancreatic cells
    • C12N5/0678—Stem cells; Progenitor cells; Precursor cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/34—Materials or treatment for tissue regeneration for soft tissue reconstruction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2202/00—Special media to be introduced, removed or treated
    • A61M2202/04—Liquids
    • A61M2202/0413—Blood
    • A61M2202/0462—Placental blood, umbilical cord blood
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2210/00—Anatomical parts of the body
    • A61M2210/14—Female reproductive, genital organs
    • A61M2210/1433—Uterus
    • A61M2210/1458—Placenta
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2210/00—Anatomical parts of the body
    • A61M2210/14—Female reproductive, genital organs
    • A61M2210/1433—Uterus
    • A61M2210/1466—Umbilical cord
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Abstract

Células madre placentarias aisladas en una cantidad terapéuticamente eficaz para uso en un método de tratamiento de un individuo que ha tenido isquemia, en donde dichas células madre placentarias son fibroblastoides y se adhieren a plástico de cultivo de tejidos, en donde dichas células madre placentarias se pueden obtener por perfusión de una placenta humana con una solución de perfusión, en donde dicha placenta ha sido exanguinada y perfundidapara retirar la sangre residual; y en donde dichas células madre placentarias: (a) no expresan antígenos MHC de clase 2: (b) son CD34- y CD38-: (c) son OCT-4+ y ABC-p+; (d) son SSEA3-, SSEA4-, OCT-4+ y ABC-p+; o (e) son CD10+, CD29+, CD34-, CD38-, CD44+, CD45-, CD54+, CD90+, SH2+, SH3+, SH4+, SSEA3-, SSEA4-, OCT-4+, y ABC-p+.

Description

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mantenimiento (BioWhittaker) disponibles en el mercado utilizados de acuerdo con las instrucciones de los fabricantes.
6.2.4. Aislamiento de células madre de tipo embrionario placentarias
El examen microscópico de las células adherentes en los matraces de cultivo reveló tipos de células morfológicamente diferentes. Se observaron células con forma de huso, células redondas con grandes núcleos y numerosas vacuolas pequeñas perinucleares, y células con forma de estrella con numerosas proyecciones (a través de una de las cuales las células con forma de estrella se anclaban al matraz) que se adherían a los matraces de cultivo. Aunque no se realizaron intentos para caracterizar adicionalmente estas células adherentes, se observaron células similares en el cultivo de médula ósea, sangre de cordón y periférica, y por lo tanto se consideró que no eran de tipo célula madre en la naturaleza. Las células fibroblastoides, que aparecían al final como agrupaciones, eran candidatas a ser MSC (células madre mesenquimales) y se aislaron mediante tripsinización diferencial y se subcultivaron en matraces secundarios. La microscopía de fase de las células redondeadas, tras la tripsinización, reveló que las células estaban muy granuladas; eran indistinguibles de las MSC derivadas de la médula ósea producidas en el laboratorio o adquiridas de BioWhittaker. Cuando se subcultivaron, las células madre de tipo embrionario derivadas de placenta, al contrario que en su fase más temprana, adheridas en horas, adoptaron una forma fibroblastoide característica, y formaron un patrón de crecimiento idéntico al de las MSC derivadas de médula ósea. Durante el subcultivo y la realimentación, por otra parte, las células mononucleares unidas laxamente se lavaron y los cultivos permanecieron homogéneos y desprovistos de cualquier contaminante de célula no fibroblastoide visible.
6.2.5. Resultados
La expresión de CD-34, CD-38, y otros marcadores de la superficie asociados a células madre sobre células mononucleares purificadas en la fracción temprana y tardía se evaluó mediante citometría de flujo. Las células clasificadas, recuperadas se lavaron en PBS y a continuación se sometieron a doble tinción con anti-CD34ficoeritrina y anti-CD38-isotiocianato de fluoresceína (Becton Dickinson, Mountain View, CA).
El aislamiento celular se logró utilizando la separación de células magnéticamente, tal como por ejemplo, Auto Macs (Miltenyi). Preferiblemente, se realiza primero el aislamiento de las células CD34+.
6.3 Ejemplo 3: medio de perfusión
El siguiente ejemplo proporciona una fórmula de la solución de perfusato preferida para el cultivo de placentas aisladas
Agente químico
Fuente Concentración de la solución de partida Concentración Final 500 ml
DMEM-LG
GibcoBRL11885-084 imagen21 imagen22 300 ml
MCDB201
Sigma M-6770 Disuelto en H2O pH a 7,2. filtro 200 ml
FCS
Hyclone 100% 2% 10 ml
ITS
Sigma I-3146 o GibcoBRL41400-045 100x 1x 5 ml
Pen y Estrep
GibcoBRL15140-122 100x 1x 5 ml
LA+BSA
Sigma+GibcoBRL BSA 100 x (1 µg/ml de LA 10 ng/ml de LA 5 ml
Dexametasona
Sigma D-2915 0,25 mM en H2O 0,05 µM 100 µl
Ácido Lascórbico
Sigma A-8960 1000 x (100 mM) 1 x (0,1 mM) 500 µl
PDGF (50 µg)
R&D 220BD 10 µg/ml en HCl 4 mM + BSA al 0,1% 10 ng/ml 500 µl
EGF (200 µg)
Sigma E-9644 10 µg/ml en HAc 10 mM + BSA al 0,1% 10 ng/ml 500 µl
La composición anterior es un perfusato que se puede utilizar a una variedad de temperaturas para perfundir la placenta. Se debe observar que se pueden utilizar en el perfusato o el medio de cultivo componentes adicionales tales como antibióticos, anticoagulantes y otros factores de crecimiento.
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hematopoyéticas enriquecidas en células CD34+ que son CD38-.
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La composición del punto 53, que comprende además sangre del cordón que tiene células que son CD34+ y CD38+.
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Una composición adecuada para el trasplante de médula ósea que comprende una población de células madre hematopoyéticas enriquecidas en células que son CD34-y CD38.
59.
La composición del punto 54, que comprende además sangre del cordón que tiene células que son CD34+ y CD38+.
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Claims (1)

  1. imagen1
ES10182243.5T 2001-02-14 2002-02-13 Placenta post-parto de mamíferos, su uso y células madre placentarias de la misma Expired - Lifetime ES2558626T3 (es)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US26856001P 2001-02-14 2001-02-14
US268560P 2001-02-14
US4942 2001-12-05
US10/004,942 US7045148B2 (en) 2000-12-06 2001-12-05 Method of collecting placental stem cells

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ES2558626T3 true ES2558626T3 (es) 2016-02-05

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Application Number Title Priority Date Filing Date
ES10182485T Expired - Lifetime ES2432493T3 (es) 2001-02-14 2002-02-13 Placenta de mamífero post-parto, su uso y células madre de la misma
ES02720974.1T Expired - Lifetime ES2545899T3 (es) 2001-02-14 2002-02-13 Placenta post-parto de mamíferos, su uso y células madre placentarias de la misma
ES10182243.5T Expired - Lifetime ES2558626T3 (es) 2001-02-14 2002-02-13 Placenta post-parto de mamíferos, su uso y células madre placentarias de la misma
ES10182303.7T Expired - Lifetime ES2549161T3 (es) 2001-02-14 2002-02-13 Placenta post-parto de mamífero, su uso y células madre placentarias de la misma
ES02709498.6T Expired - Lifetime ES2444548T3 (es) 2001-02-14 2002-02-13 Renovación y repoblación de tejidos y órganos cadavéricos descelularizados por células madre
ES10183252.5T Expired - Lifetime ES2629155T3 (es) 2001-02-14 2002-02-13 Matrices de tejido que comprenden células madre placentarias, y métodos para su preparación
ES10182362.3T Expired - Lifetime ES2522526T3 (es) 2001-02-14 2002-02-13 Placenta post-parto de mamíferos, su uso y células troncales placentarias de la misma
ES10183301.0T Expired - Lifetime ES2559031T3 (es) 2001-02-14 2002-02-13 Renovación y repoblación de tejidos descelularizados y órganos cadavéricos por células madre
ES10182433.2T Expired - Lifetime ES2548591T3 (es) 2001-02-14 2002-02-13 Placenta post-parto de mamífero, su uso y células madre placentarias de la misma

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ES10182485T Expired - Lifetime ES2432493T3 (es) 2001-02-14 2002-02-13 Placenta de mamífero post-parto, su uso y células madre de la misma
ES02720974.1T Expired - Lifetime ES2545899T3 (es) 2001-02-14 2002-02-13 Placenta post-parto de mamíferos, su uso y células madre placentarias de la misma

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