Cell Culture Flask

Content
1. What is cell culture flask?
2. Suspension cell culture flask
3. Types of cell culture flask
4. Use of cell culture flask
    4.1 How to better digest cells in cell culture flasks?
    4.2 How to change the solution in the cell culture flask?
5. How to buy cell culture flask?

Cell culture is a very important experimental technique and has become an indispensable research modality in the fields of biopharmaceuticals, life sciences, and clinical transplantation. Cell culture must be done with the help of cell consumables to achieve the necessary conditions for cell growth, and cell culture flask is one of the most common laboratory tools.

Cell culture flasks allow for the successful growth and proliferation of microbial, insect, or mammalian cells. The in vitro culture of cells and tissues has become a part of life research and practice. A wide range of cell types is cultured, from viruses to bacteria and fungi, and from human cells to animal and plant cells. Some cells and tissues can be grown in suspension, but a significant proportion of mammalian cells require surface apposition. Therefore, the culture flasks, plates, and dishes used to provide in vitro cell culture environments are not only transparent and non-toxic but also sterile. They also need to be surface-modified to enable them to adhere, divide and grow.

Since entering the 21st century, biology has entered a breakthrough stage of development, and the position of the life sciences in the natural disciplines has been revolutionized. Among them, molecular biology, genetics, cell biology, developmental biology, neurology, ecology, space life sciences, and other disciplines will gradually develop into leading disciplines.

Molecular biology (including molecular genetics) in the mainstream of the life sciences, as well as its great role in promoting the development of the life sciences as a whole is indisputable. The cell is the basic structural and functional unit of life activity, cell biology as the basic discipline of biological science status also needs to be given attention. Speaking of cell culture, according to the current classification of cells are mainly divided into: suspension cells and wall cells. The culture of suspension cells is now the mainstream and efficient cell culture in the life sciences. The containers required for suspension cell culture experiments have also become a basic tool in the work of scientists.
At present, there are three kinds of common suspension cell culture vessels as follows.

In 1861, the national scientist Erlenmeyer, in order to ensure the heat stability of the sample reaction flask, improved the heating vessel and invented the conical flask. The excellence of the conical flask is reflected in its special shape. Compared to a round beaker, this special shape increases the stability of the instrument and makes it less likely to be knocked over. In addition, during the shaking process, the "neck" of the conical flask makes the liquid inside the flask less likely to splash out, so that the culture solution will not splash out of the vessel during the suspension cell culture process, thus ensuring the stability of the reaction system. Nowadays, typical cultures of CHO, SF9, NK cells, etc. are grown in shake flasks. Theoretically, in order to ensure a good amount of dissolved oxygen, the volume of the shake flask culture is about 10% of the vessel volume, but it may be increased in practice.

In mammalian and insect cell cultures, where cells exhibit high viability and protein expression, ANTITECK bottles of all models show a significant increase in cell viability and activity in cell culture. Thanks to the unique low-shear spoiler of the cell culture flask, the aeration efficiency is increased, so that the filling volume can reach 60% of the total volume. Other types of shake flasks can hold only 30% of the total volume.

The cell culture flask enables healthy cell growth, resulting in increased expression of target products such as membrane proteins, secreted proteins, and monoclonal antibodies; increasing insect cell protein expression by 200%. For some stable cells and transient expression cells, protein expression can be increased by 300%.

Using cell culture flasks, the ratio of the volume of filling liquid is increased and the expression per unit volume is increased. The consumption of culture flasks and culture medium can be saved substantially. Cell culture flasks with high reproducibility, scalable sub-cell growth from batch to batch, and high consistency in yield.

The culture flask, named after the French biologist Auguste Fernbach, is actually an improvement on the shake flask. The bottom area of the shake flask has been enlarged, thus increasing the area of oxygen exchange between the culture medium and the air. Both flat and threaded openings are available, so a breathable lid can be selected. The large opening reduces the difficulty of transferring the medium. The baffle at the bottom increases the flow of liquid when the bottle is shaken to increase the probability of contact between the liquid and oxygen. The Bach bottle increases the product yield more than a regular shake bottle.

Cell culture is a method to simulate the in vitro environment in order to survive, grow, reproduce and maintain major structures and functions. A variety of cell culture consumables are used to culture cells, of which cell culture flasks are one of the more heavily used.

The shape of the cell culture flask is designed to be square with a wide neck, which is intended to facilitate the harvesting of cells. The frosted design is usually used on the side of the bottle to facilitate the operator's record. In terms of specifications, this consumable is commonly available in 25cm2, 75cm2, 175cm2, 225cm2, etc. The specifications we usually refer to are the capacity of the culture medium that the bottle can hold. According to the different specifications, there are different application scenarios.

The small-size bottles of 25 cm2 and 75 cm2 are mainly used for cell recovery and small-scale expansion, mainly in the pre-culture phase. It can also be used as a storage container for a variety of red blood cells. The larger size bottles of 175 cm2 and 225 cm2 are mainly used for medium-scale cell culture or eukaryotic protein expression, etc. The larger size bottles of 175 cm2 and 225 cm2 are mainly used for medium-scale cell culture or eukaryotic protein expression. Protein expression is a molecular biology technique that uses bacteria, yeast, insect cells, mammalian cells, or plant cells to express exogenous gene proteins, and is one of the important components of genetic engineering technology.

Cell culture flasks are mainly used for the culture of walled cells. Although different specifications differ in their use, they all have to meet the requirements for cell growth, such as no DNA enzymes, no RNA enzymes, no endotoxins, no animal sources, surface TC treatment, etc.

Cell culture flask is one of the most used cell culture consumables when culturing cells. Cultivating cells is a very rigorous task, and it requires the right conditions for better growth. The harvesting of cells is involved when growth reaches a certain level. When using cell culture flasks, the following four steps can be followed.

a. Pour off the old medium and add a small amount of new medium to wash 1-2 times, the purpose of doing so is to wash away as many dead cells floating around.

b. Add a small amount of new medium and blow it directly once, this time to blow down the cells that are not firmly attached to the wall. Wash again with medium and mix the suspension twice to pass into a new cell culture flask.

c. Aspirate the remaining liquid in the bottle, add about 0.3 ml of trypsin to wash it once, aspirate it off and discard it, and then add about 1 ml of trypsin to digest it. When the gap between cells and cells is obvious, immediately aspirate and discard the trypsin, add a new medium and start to blow, blow 2-3 times and then aspirate the suspension in a test tube or new bottle for temporary storage. Wash with 2ml of new medium and mix with the previous one.

d. Continue to digest the remaining firmly adherent cells with fresh trypsin added to the flask. Continue to observe microscopically, and when the remaining cell gaps are evident and the cells are independently open, aspirate off the trypsin, add a new medium and blow and pass the suspension into a new bottle for culture.

The above steps will result in a better quality of cells digested from the cell culture flasks and better growth status during the later passages.

In the process of culturing cells, the fluid exchange is a very routine operation, which is mainly to remove various metabolic wastes produced by cells during growth, replenish fresh culture medium and promote cell growth.

To carry out the following steps, an alcohol spray bottle containing 75% medical sterile alcohol, PBS buffer, serum-containing medium, a Pap syringe, a pipette, and a waste jar is required.

a. Put the equipment into the ultra-clean table and turn on the UV irradiation for sterilization. If the cultured cells are temperature sensitive, you can put the serum-containing media bottles into a 37°C water bath to make the media temperature at 37°C, and then transfer them to the ultra-clean table for UV sterilization. In addition, the cell culture room also needs UV irradiation for about half an hour.

b. Remove the cell culture bottle from the incubator, disinfect the bottle surface with a 75% alcohol spray bottle, transfer to the ultra-clean table, open the lid, gently aspirate out the old culture medium, and add fresh serum-containing medium with a pipette. Note that the medium must not be added from the adnexal side of the cells, but from the side wall, with gentle movements so as not to wash off the cells.

c. After adding the medium, close the lid and mark the bottle with information such as time of fluid change, cell type, and operator.

d. Before putting the bottles into the incubator, you should spray the surface of the bottles with alcohol again, and then you should remember to organize the operating table, wipe the ultra-clean table surface with alcohol, and clean up the waste liquid and garbage.

When performing the liquid exchange operation, pay attention to asepsis throughout the process. Operators should wear sterilized clothing, sterilized gloves, and masks, and must be careful when their hands touch the mouth of the cell culture bottle to avoid contamination.

If you are interested in our cell culture flask or have any questions, please write an e-mail to [email protected], we will reply to you as soon as possible.