CN204918617U - a cell culture flask - Google Patents

Abstract

The utility model discloses a cell culture bottle applicable to the field of biopharmaceuticals, which is composed of a bottle body made of glass that can be repeatedly high-pressured, and is connected with an internal thread bottle cap that can be repeatedly high-pressured through an external threaded bottle mouth. Appropriately open the hole, install the hydrophobic microporous filter membrane in the bottle cap, and press and fix it with the same open-hole silica gel gasket. Its technical effect is that the hydrophobic microporous filter membrane can be replaced regularly, the glass bottle body can be repeatedly cleaned, and all parts can be repeatedly autoclaved, so that the cell culture bottle can achieve good air permeability under sterile conditions, and can avoid The culture medium is over-evaporated, and the cell culture bottle can be used repeatedly, which greatly reduces the cost of suspension cell culture, and provides an economical cell culture mode for the research and development of biopharmaceuticals prepared by suspension cell culture.

Description

a cell culture flask

technical field

The utility model relates to a cell culture bottle in the field of biopharmaceutical equipment.

Background technique

Cell culture flasks are important consumables for cell culture and drug production in the biopharmaceutical industry. Especially in the field of recombinant protein production, the requirements for sterility and air permeability of cell culture flasks are quite high, so they are widely used by researchers in the pharmaceutical industry and medical device industry. favor. Cell culture flasks can be divided into glass cell culture flasks and plastic cell culture flasks according to the material; according to the shape of the bottom of the culture flask, they can be divided into round, square, rectangular, triangular, etc. According to the volume of the cell culture bottle, it can have various specifications such as 5-1000ml. According to the cell growth mode, it can be divided into adherent cell culture flask and suspension cell culture flask.

The rapid development of the biopharmaceutical industry has made mammalian cell suspension culture the mainstream recombinant protein production technology. Mammalian cell suspension culture usually uses disposable plastic cell culture bottles, because this kind of culture bottles are safe and mature, and the bottle caps are available with sealing caps, vent caps, etc.; however, the cost of using disposable culture bottles is also very high , not only brings a heavy economic burden to researchers, but also causes serious waste of resources in the process of research and production; while glass cell culture bottles can be repeatedly autoclaved, greatly reducing the cost of use, but at present, glass cell culture bottles only have spiral seals Cover, the sealing cover needs to be unscrewed during cell culture to ensure the flow rate and pH stability of the cell culture. Such an operation is not only inconvenient, but also greatly increases the risk of contamination during the culture process.

Therefore, there is a need for a reusable glass cell culture bottle, which has a sterile air-permeable bottle cap at the same time, so as to meet the need for gas exchange under aseptic conditions during the culture process.

Utility model content

The purpose of the utility model is to overcome the deficiencies of the prior art and provide a cell culture bottle that can be used repeatedly, which can ensure gas exchange under sterile conditions during the cell culture process.

One solution to achieve the above object is a cell culture bottle, which includes a bottle body and a bottle cap, and the bottle cap is connected to the bottle body through a screw.

The bottle cap is made of high-density polyethylene (HighDensityPolyethylene, referred to as "HDPE"). A punching machine is used to open a ring hole in the center of the bottle cap, and a disc-shaped hydrophobic microporous filter with the same diameter as the inner diameter of the bottle cap is installed in the bottle cap. Membrane to provide sterile gas exchange for the cells being cultured in the cell culture flask and reduce the risk of contamination of the cells in the cell culture flask. The hydrophobic microporous filter membrane can be purchased from the market, and the filter pore size is usually 0.1-0.45 μm, preferably 0.22 μm; the filter membrane is a hydrophobic filter membrane commonly used in open cell culture, and is especially suitable for cell culture in a _CO2 incubator. Culture, especially for experiments that require long-term cell culture.

Further, in order to fix the microporous filter membrane installed in the bottle cap, use a silicone gasket with the same diameter as the inner diameter of the bottle cap to punch out an annular hole with the same pore size for pressing and fixing the filter membrane; the silicone gasket can be used with flat tweezers Take it out and put it back for easy filter replacement.

Further, in order to ensure air permeability under sterile conditions and prevent excessive evaporation of the medium, it is necessary to limit the opening size of the bottle cap according to the volume of the bottle body. Generally, the diameter of the opening is 1/10-1/3 of the diameter of the bottle cap. Preferably 1/5.

Further, the hydrophobic microporous filter membrane installed in the bottle cap can be replaced with a new filter membrane after repeated high pressure 10-20 times to ensure gas exchange under sterile conditions.

Further, the bottle body is made of glass;

Further, the bottle body is made of borosilicate glass.

Still further, the bottle mouth is an external thread; the bottle cap is correspondingly an internal thread.

Still further, the bottle body can be silanized to reduce cell adsorption.

A technical scheme of a cell culture bottle of the present invention is adopted, that is, a bottle body made of glass that can be repeatedly high pressure is connected with an internal thread bottle cap that can be repeatedly high pressure through an external threaded bottle mouth, and a hole is appropriately opened in the middle of the bottle cap. A hydrophobic microporous filter membrane with a pore size of preferably 0.22 μm is installed in the bottle cap, and it is compressed and fixed with a similarly opened silica gel gasket. Therefore, the cell culture bottle can achieve good gas permeability under sterile conditions, and can avoid excessive evaporation of the medium, and the entire cell culture bottle can be used repeatedly after thorough cleaning and autoclaving, greatly reducing the cost of suspension cell culture , providing an economical cell culture model for the development and development of biopharmaceuticals prepared using suspension cell culture.

Description of drawings

Fig. 1 is the front view of a kind of cell culture bottle of the present utility model.

Fig. 2 is a component diagram of a cell culture flask of the present invention. in

1: bottle body; 2: threaded bottle mouth; 3: hole threaded bottle cap

Fig. 3 is a top view of a bottle cap of a cell culture bottle of the present invention.

3: Open-hole threaded cap; 4: Ring-opening sealing gasket; 5: Hydrophobic microporous filter membrane.

Fig. 4 is a graph of cell viability and cell density of CHO cultured in a cell culture flask of the present invention. Wherein, a is the cultured cells in the commercial disposable cell culture bottle; b is the cultured cells in the cell culture bottle of the present utility model after cleaning and disinfecting once; c is the cultured cells in the cell culture bottle of the present utility model after cleaning and disinfecting for 10 times.

Detailed ways:

Please refer to Fig. 1, Fig. 2 and Fig. 3. In order to better explain the technical solution of the present utility model, the inventor of the present utility model will describe in detail through specific embodiments and in conjunction with the accompanying drawings.

Please refer to Fig. 1 and Fig. 2, a kind of cell culture bottle of the present utility model (Fig. 1), comprises bottle body 1 and bottle mouth 2 of external thread, special internal thread bottle cap 3, bottle cap 3 connects bottle mouth 2 through thread Tight junctions (Figure 2), construct sterile conditions to prevent contamination of cells cultured in cell culture flasks.

There is an annular opening in the center of the bottle cap 3 (Figure 3), and a hydrophobic microporous filter membrane is installed inside, and it is compressed and fixed with a silica gel gasket with the same opening. The purpose of this design is to not only ensure the air permeability of the bottle cap, but also create sterile conditions and prevent the medium from evaporating. At the same time, the cell culture bottle can be reusable by cleaning and replacing the hydrophobic microporous filter membrane and then autoclaving. The purpose of use is to greatly reduce the cost of suspension cell culture and reduce the resource consumption of biopharmaceutical research and development.

In this example, cell culture bottles of different volumes are equipped with bottle caps of different diameters (Figure ₂ ). nourish.

In this example, the CHO cells were cultured in a CO ₂ incubator using the cleaned and autoclaved cell culture flasks. The temperature was controlled at 37° C., the CO ₂ concentration was 5%, and the shaker speed was 120 rpm. The cell density was 3×10 ⁵ cells/ml; samples were taken every 24 hours for cell counting and viability determination. The results are shown in Figure 4. The cell viability was maintained above 95%, and the cell growth curve was also the same as that of commercial products. Sex cell culture bottle is quite, confirms that the design of described cell culture bottle is reasonable and feasible.

Claims (9)

1. a cell culture bottle, comprising bottle body, bottleneck and bottle cap, described bottle cap is connected with described bottle body by the screw thread of bottleneck, it is characterized in that, described bottle cap is perforated design, perforate diameter It is 1/10-1/3 of the diameter of the bottle cap, and a microporous filter membrane with the same size as the bottle cap is installed inside the bottle cap. The bottle body is made of glass, and the diameter-to-height ratio of the bottle body is 2:1-1 :1. 2. The cell culture bottle according to claim 1, characterized in that the material of the bottle cap is high-density polyethylene, which is designed with an annular hole in the center. 3. The cell culture bottle according to claim 1, characterized in that the diameter of the opening is 1/5 of the diameter of the bottle cap. 4. The cell culture flask according to claim 1, characterized in that, the filter aperture of the microporous membrane is 0.1-0.45 μm. 5. The cell culture flask according to claim 4, characterized in that the filter aperture of the microporous membrane is 0.22 μm. 6. The cell culture flask according to claim 4 or 5, characterized in that, the microporous filter membrane is compressed and fixed with a similarly perforated silica gel sealing ring. 7. The cell culture bottle according to claim 6, characterized in that 1-3 layers of microporous filter membranes are installed and the filter membranes can be replaced at will. 8. The cell culture bottle according to claim 1, characterized in that the bottle body is made of borosilicate glass. 9. The cell culture bottle according to claim 1, characterized in that: the diameter-to-height ratio of the bottle body is 3:2.