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Monoclonal enhanced culture medium and method for improving HepG2 cell clone formation rate

A cell culture and culture medium technology, applied in the field of HepG2 cell cloning, can solve the problem of low monoclonal formation rate of HepG2 cells, and achieve the effects of improving the monoclonal formation rate, fast growth rate, and increased clone formation rate.

Active Publication Date: 2022-05-31
广州源井生物科技有限公司
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Problems solved by technology

[0004] In view of the problems raised by the background technology, the purpose of the present invention is to propose a monoclonal enhanced medium, and the monoclonal formation rate of HepG2 cells is significantly improved by using the monoclonal enhanced medium. This invention solves the problem of HepG2 cells under conventional culture conditions. The problem of low monoclonal formation rate of
[0005] Another object of the present invention is to propose a method for improving the clone formation rate of HepG2 cells, which can significantly improve the clone formation rate of HepG2 cells and solve the problem that the clone formation rate of HepG2 cells is low under conventional culture conditions

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  • Monoclonal enhanced culture medium and method for improving HepG2 cell clone formation rate

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Embodiment 1-7

[0070]A method for improving the clone formation rate of HepG2 cells, comprising the following steps:

[0071] (1) Primary culture and subculture of HepG2 cells are carried out in the following order;

[0072] Specifically, in step (1), the operation steps for primary culture of HepG2 cells are as follows:

[0073] (1.1.1) Preparation materials: DMEM complete culture medium, T25 culture bottle, 15mL centrifuge tube, 10mL serological pipette, waste liquid tank, alcohol cotton, etc.;

[0074] Preparation: Preheat the complete culture solution in a 37°C water bath for 30 minutes, then remove the frozen cells from the liquid nitrogen, transfer them to a -80°C refrigerator, and let the residual liquid nitrogen evaporate for 5 minutes;

[0075] (1.1.2) Use a straw to draw 6mL of complete culture solution into a 15mL centrifuge tube in the ultra-clean bench;

[0076] (1.1.3) Take the HepG2 cells out of the -80°C refrigerator and temporarily place them in dry ice, and shake them sli...

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Abstract

The invention discloses a monoclonal enhancement culture medium and method for improving the HepG2 cell cloning formation rate, and relates to the technical field of HepG2 cell cloning. The monoclonal enhancement culture medium comprises the following components: an RPMI-1640 culture medium, a DMEM culture medium, fetal calf serum, a hepatocyte growth factor and an insulin-like growth factor-1; the method for improving the HepG2 cell clone formation rate comprises the following steps: sequentially carrying out primary culture and subculture on HepG2 cells, preparing monoclone by adopting a limited dilution method, and after culturing for 24-48 hours, replacing a basal culture medium with a monoclonal enhancement culture medium, and continuously culturing to form the monoclone. By using the monoclonal enhanced culture medium provided by the invention, the monoclonal formation rate of the HepG2 cells is remarkably improved, and the problem that the monoclonal formation rate of the HepG2 cells is relatively low under conventional culture conditions is solved.

Description

technical field [0001] The invention relates to the technical field of HepG2 cell cloning, in particular to a monoclonal enhancement medium and a method for improving the formation rate of HepG2 cell clones. Background technique [0002] HepG2 cells were isolated from the liver tissue of a 15-year-old male with well-differentiated hepatocellular carcinoma. The biotransformation metabolic enzymes contained in HepG2 cells are homologous to those of normal human liver parenchymal cells, and are often used in drug metabolism and liver cancer. Toxicity research, specifically gene editing through ZFN, TALEN and CRISPR / Cas9 technology, so as to establish gene knockout, knockin, point mutation and other cell lines are commonly used research methods for HepG2 cells, drug metabolism and liver toxicity of HepG2 cells The research made a huge achievement for the later research on the treatment of liver cancer. [0003] With the in-depth study of drug metabolism and liver toxicity, the ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/09
CPCC12N5/067C12N5/0693C12N2501/105C12N2501/12C12N2500/32
Inventor 黄秋凤郑虹李榕贞
Owner 广州源井生物科技有限公司
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