Application of cell culture flasks in the passage of mouse fibroblasts

 Cell culture flasks are a type of cell culture consumables that are widely used in the market, and are mainly suitable for adherent cell culture. Among them, this consumable is also used in the passage of mouse fibroblasts.

Mouse fibroblasts are special cells used for non-medical purposes such as scientific research or industrial applications. Fibroblasts have vigorous functions and weak basophilic cytoplasm. They have obvious protein synthesis and secretion activities. Under certain conditions, they Can achieve mutual transformation with fiber cells. After receiving the fibroblasts, they need to be stabilized in the incubator for about 4 hours and then cultured or passaged according to the cell density, changing the medium. The passaging operation follows the following steps:

1. Aspirate the medium in the original cell culture flask, rinse the cells twice with PBS buffer, and add 1~2 ml of 0.25% EDTA for trypsin digestion (pay attention to the digestion time, usually controlled within 1~2min).

Application of cell culture flasks in the passage of mouse fibroblasts

2. Observe the digestion under the microscope. When the cell edge shrinks and adheres to the wall (digestion is not recommended until the cells float), remove the trypsin, add 6~8ml of complete medium, gently blow the cell layer, and try to blow off and blown away the cell layer.

3. Take out part of the cell suspension and transfer it to a new cell culture flask, add appropriate complete medium, and cultivate in an incubator.

4. Pay attention to the change of the pH value of the medium, change the medium regularly, and repeat the subculture or freeze storage when the cell density reaches 70-80%.

The main role of cell culture flasks in cell passaging is to store cells and provide an adherent growth environment for cells. It should be noted that if the cell wall is broken or leaking after receiving the cells, it means that the cells have been contaminated, and the supplier should be contacted in time.

评论

发表评论

此博客中的热门博文

Two commonly used materials for carboys

图片
  In biological experiments, the processing and preservation of some liquids are often involved.   Carboys   are ideal for storing sterile liquids, distributing solutions and culture media. The commonly used specifications are mainly 10L and 20L. The material of this container are two main types: PP (polypropylene) and PC (polycarbonate). Polypropylene: It is a polymer formed by the addition polymerization reaction of propylene. The operating temperature range is -30~140℃. It can resist the corrosion of acids, alkalis, salt liquids and various organic solvents below 80℃, and can decompose under high temperature and oxidation. Polypropylene is a thermoplastic synthetic resin with excellent properties. It is a colorless, translucent thermoplastic lightweight general-purpose plastic with chemical resistance, heat resistance, electrical insulation, high-strength mechanical properties and good high-wear resistance processing properties.   Polycarbonate: The liquid storage...
阅读全文

What characteristics do the culture media bottles need to meet?

图片
  The media is the basic substance that provides cell nutrition and promotes cell proliferation, and is also the living environment for cell growth and reproduction. There are many types of medium, the raw materials for preparation are different, and the requirements for use are different. What characteristics should the   media bottles   meet to ensure that the medium is well preserved? The general medium is easily contaminated or decomposed by bacteria after being heated and absorbing moisture. Therefore, the general medium must be kept in a cool and moisture-proof place. For some media (such as tissue culture medium) that need to be strictly sterilized, they must be stored in a refrigerator at 3-6 °C for a long time. To ensure good storage of the medium, the media bottle should meet the following characteristics: 1. Sterility: The medium bottle must be aseptically treated to ensure that the medium is not contaminated. The general sterilization method will choose electr...
阅读全文

Cause Analysis of Mycoplasma Contamination in Cell Factory

图片
  Mycoplasma contamination of cells is a very troublesome problem when culturing cells, especially in the use of large-scale culture vessels such as   cell factory . Once mycoplasma contamination occurs, the loss will be even greater. So, what is the cause of mycoplasma contamination? Mycoplasma is a prokaryotic organism with a size of only 0.2-0.3 um, no cell wall, and can pass through a general filter membrane (0.22-0.45 um). During cell culture, the incidence of mycoplasma infection reaches 63%, so during cell culture Contamination by mycoplasma is a worldwide problem. When cells (especially passage cells) are contaminated by mycoplasma, the expression of DNA, RNA and protein in the cells will change, but the growth rate of cells generally does not have a significant impact, so the contamination of cells by mycoplasma is generally difficult to detect. In most cases, the cytopathic changes are slight or insignificant, and subtle changes can also be alleviated by subculture a...
阅读全文