Infection enhancing culture medium and method for improving cell lentivirus infection rate

Abstract

The invention discloses an infection enhancing culture medium and method for improving the cell lentivirus infection rate, and relates to the field of MDA-MB-231 cell transfection. The enhanced infection culture medium is used for improving the lentivirus infection rate of the MDA-MB-231 cell, and the enhanced infection culture medium is prepared from the following components: a Leibovitz's L-15 culture medium, an RPMI-1640 culture medium, fetal calf serum, non-essential amino acid and an epidermal growth factor; according to the volume percentage of the total raw materials, the addition amount of the fetal calf serum is 10%, and the addition amount of the non-essential amino acid is 1%; the addition amount of the epidermal growth factor is 2-3ng/ml. By adopting the enhanced infection culture medium disclosed by the invention, the MDA-MB-231 cells can be normally cultured in the conventional environment of 37 DEG C and 5% CO2, and the infection efficiency of the lentivirus on the MDA-MB-231 cells is more remarkably improved, so that the gene editing efficiency of the MDA-MB-231 cells becomes higher, and the problem that the lentivirus infection rate of the MDA-MB-231 cells is low at present is solved.

Description

technical field [0001] The invention relates to the field of MDA-MB-231 cell transfection, in particular to an enhanced infection medium and method for increasing the infection rate of cell lentivirus. Background technique [0002] MDA-MB-231 (human breast cancer cells) are human breast cancer cells isolated from the pleural effusion of a 51-year-old female breast cancer patient, MDA-MB-231 is a highly invasive and poorly differentiated triple-negative breast cancer (TNBC) cell line, triple negative refers to lack of estrogen receptor (ER) and progesterone receptor (PR) expression, and HER2 (human epidermal growth factor receptor 2) overexpression, similar to other invasive cancer cell lines, The invasiveness of MDA-MB-231 cells is mediated by proteolytic degradation of the extracellular matrix. [0003] MDA-MB-231 is widely used in research on triple-negative breast cancer. Gene editing through ZFN, TALEN, and CRISPR / Cas9 technologies to establish gene knockout, knockin, a...

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Problems solved by technology

[0003] MDA-MB-231 is widely used in research on triple-negative breast cancer. Gene editing through ZFN, TALEN, and CRISPR / Cas9 technologies to establish gene knockout, knockin, and point mutation cell lines has become the mainstream of research. Means, MDA-MB-231 cell conventional transfection method is the lentivirus method, although it can infect, but the efficiency is relatively low, only about 30%, and it needs to be screened by introducing additional resistance genes to increase the infection rate, but through drug The stable cells enriched by screening, the cell state is poor, and the positive rate of knockout in pool cells and monoclonal identification is relatively low, which restricts the gene editing efficiency of MDA-MB-231 cells

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