CN209890645U - Culture vessel for pit bottom adherent cells - Google Patents

Abstract

The utility model discloses a pit bottom adherence cell culture container, the cell adhesion surface of adherence cell culture container bottom is equipped with numb point form pit, numb point form pit is at cell adhesion surface evenly distributed, and the density of numb point form pit is 10 ~ 100/cm². The utility model provides a pit bottom adherence cell culture container, simple structure, convenient operation as long as once inoculate the cell, can scrape repeatedly and get the cell, need not inoculate the cell again, and is very convenient, saves time, laborsaving, economizes reagent.

Description

Culture vessel for pit bottom adherent cells

Technical Field

The utility model relates to a cell culture container technical field, concretely relates to pit bottom adherence cell culture container.

Background

All in vitro cultured cells, including primary cultures and various cell lines, need to be subcultured when the cells grow to a certain density. Suspension culture cell passage is relatively easy, and adherent cell's passage is relatively complicated, and adherent cell grows into compact monolayer after the blake bottle, and the space of continuing to grow is not enough, and the nutrient composition in the culture medium also consumes more, and metabolic waste also has more to pile up simultaneously, needs the bottle culture of branch, carries out passage expansion to the cell.

For cells with less tight adherence, such as 293T cells, they can be blown off directly and then bottled. For cells with tight adherence, such as CHO cells, digestion with 0.25% pancreatin solution (with or without EDTA) and then blowing to disperse the bottle are required, and the general process is as follows (the following operations are carried out according to the requirements of aseptic operation):

discarding the original culture solution in the cell bottle growing into the compact monolayer cells; adding 0.25% pancreatin solution (with or without EDTA) in a volume of 0.5ml or more depending on the size of the cell vial to allow sufficient infiltration; generally, the digestion time is about 1 to 3 minutes, a semitransparent cell layer on the wall of a bottle is observed by naked eyes, when a fine pinhole gap appears, a pancreatin solution is discarded, and a proper amount of cell culture solution is added to stop digestion; the cells are blown off, washed by PBS and centrifuged, and are divided into a plurality of bottles to be continuously cultured according to the experimental requirements, and the ratio of one to two to one to four is generally adopted.

If the aim of repeatedly harvesting cells is to digest adherent cells by using the cell digestion solution, most of the cells can be used for subsequent application, a proper amount of cells are remained as seed cells, the seed cells are remained in a culture container, and a fresh culture solution is added for continuous culture. The process of digesting cells and reseeding cultured cells is repeated repeatedly to obtain cells continuously, which is time-consuming, tedious and increases the pollution chance.

Some experiments require that cells cannot be treated by cell digestive fluid, proteins or sugar chains on the surfaces of the cells are prevented from being damaged, and only cells can be collected by cell scraping or cell shoveling. The cell adherent surface of the existing cell culture vessel is a very smooth plane, once a cell scraper or a cell shovel is applied, all cells can be scraped off, and if the cells need to be continuously cultured, the cells need to be inoculated again, which wastes time and labor.

The present invention is made to solve the above inconvenience.

SUMMERY OF THE UTILITY MODEL

The utility model aims at providing a pit bottom adherence cell culture container solves one or more among the above-mentioned prior art problem.

According to the utility model discloses a pit bottom adherence cell culture container, the cell adhesion surface of adherence cell culture container bottom is equipped with numb point form pit, numb point form pit is at cell adhesion surface evenly distributed, and the density of numb point form pit is 10 ~ 100/cm²。

The utility model provides a pit bottom adherence cell culture container, simple structure, convenient operation as long as once inoculate the cell, can scrape repeatedly and get the cell, need not inoculate the cell again, and is very convenient, saves time, laborsaving, economizes reagent.

In some embodiments, the pockmark-like dimples are cylindrical dimples, circular arc dimples, hemispherical dimples, square dimples, or conical dimples. The convenient cells will take the pit as the center and proliferate and grow to the periphery.

In some embodiments, the pit bottom adherent cell culture vessel is subjected to TC treatment. TC is fully called: the Tissue culturetreated and TC treatment means that the vessel is subjected to surface modification treatment and is suitable for culturing adherent cells.

In some embodiments, the adherent cell culture vessel is a cell culture flask, a cell culture dish, a cell culture plate, or any other cell culture equipment for culturing adherent cells, and the application range is wide, and any equipment can be applied.

In some embodiments, the adherent cell culture vessel is made of polyvinyl chloride or glass. Different materials can be selected according to actual requirements.

Drawings

Fig. 1 is a schematic structural view of a bottom cell culture surface of a pockmark bottom adherent cell culture vessel according to an embodiment of the present invention.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings.

As shown in figure 1, the pockmark bottom adherent cell culture vessel comprises pockmark-shaped pits 3 arranged on a cell attachment surface 2 at the bottom of the adherent cell culture vessel 1, wherein the pockmark-shaped pits 3 are uniformly distributed on the cell attachment surface 2, and the density of the pockmark-shaped pits 3 is 10-100/cm²The density used in this example was 20/cm²The pits 3 are cylindrical pits, arc pits, hemispherical pits, square pits or conical pits, the hemispherical pits are adopted in the embodiment, the diameter of each hemispherical pit is 50-1000 μm, the diameter of each hemispherical pit is 500 μm, the depth of each hemispherical pit is 50-800 μm, the depth of each hemispherical pit is 300 μm, and the pits 3 are dotted so that cells can conveniently proliferate and grow around the pits; the pits 3 and the flat parts of the cell attachment surfaces 2 around the pits 3 are also modified with polystyrene surfaces, and hydroxyl (OH), Carboxyl (COOH), and amino (NH and NH) groups are grafted₂) Hydrophilic groups to facilitate cell attachment.

In the above, the adherent cell culture vessel is a cell culture flask, a cell culture dish, a cell culture plate or any other cell culture equipment for culturing adherent cells, and the application range is wide, and any equipment can be used.

In the above, the adherent cell culture vessel is made of polyvinyl chloride or glass, and different materials can be selected according to actual requirements.

After the cells are scraped by a cell scraper or a cell shovel, the cells can be continuously cultured only by adding fresh cell culture solution, and the cells can be ensured to grow in an adherent manner on the cell attachment surface 2 of the adherent cell culture vessel 1.

The shapes and specifications of the cell culture bottle, the cell culture dish and the cell culture plate are the same as those of the existing commercially available cell culture bottle, cell culture dish and cell culture plate, and equipment with different specifications can be selected according to requirements; the cell culture flasks are classified into various categories according to the size of the bottom area, and the cell culture dishes are classified into various categories according to the size of the bottom area, namely, cell culture plates (generally, 6-well plates, 12-well plates, 24-well plates, 48-well plates and 96-well plates).

The using method comprises the following steps: inoculating adherent cells according to a conventional method, amplifying the adherent cells according to the conventional method until the cells are fully distributed on the whole cell culture surface, scraping the cells by using a cell scraper under an aseptic environment, scraping the cells attached to the cell culture surface, and avoiding scraping the cells in the pit-shaped pits 3; the scraped cells are taken away, fresh cell culture solution is added into the culture container again, the cells can be continuously cultured without re-inoculating the cells, the cells take the pit-shaped pits 3 as the center and proliferate and grow to the periphery, and the pit-shaped pits 3 are uniformly distributed, so the continuously cultured cells can be uniformly distributed on the whole cell culture surface after proliferating.

So, as long as once inoculate the cell, can scrape the cell repeatedly, and need not inoculate the cell again, it is very convenient, labour saving and time saving save reagent, cell culture container have pockmark form pit 3, need not to inoculate repeatedly, the cell can propagate infinitely, and the distribution is even.

The utility model provides a pit bottom adherence cell culture container, simple structure, convenient operation as long as once inoculate the cell, can scrape repeatedly and get the cell, need not inoculate the cell again, and is very convenient, labour saving and time saving province reagent.

The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, similar modifications and improvements can be made without departing from the inventive concept, and these should also be considered as within the protection scope of the present invention.

Claims (5)

1. The utility model provides a pit bottom adherence cell culture container which characterized in that, the cell adhesion surface of adherence cell culture container bottom is equipped with numb point form pit, numb point form pit and cell adhesion surface plane part around the numb point form pit is equipped with the polystyrene surface layer, is fit for the cultivation of adherence cell, the cell adhesion surface between the numb point form pit is the plane setting, numb point form pit is at cell adhesion surface evenly distributed, and the density of numb point form pit is 10 ~ 100/cm²。

2. The vessel for culturing cells adherent to the bottom of a pit according to claim 1, wherein the pits in the shape of a dot are cylindrical pits, circular arc pits, hemispherical pits, square pits or conical pits.

3. The pith bottom-adherent cell culture vessel of claim 1, wherein the pith bottom-adherent cell culture vessel is subjected to TC treatment.

4. The culture vessel of adherent cells with pith bottom of claim 2, wherein the culture vessel of adherent cells is a culture flask, a culture dish, a culture plate or any other culture equipment for culturing adherent cells.

5. The culture vessel for adherent cells with pit bottom of claim 4, wherein the culture vessel for adherent cells is made of polyvinyl chloride or glass.

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