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

A medium and infection rate technology, applied in the direction of cell culture active agent, virus, culture process, etc., can solve the problems of gene editing efficiency constraints, low knockout positive rate, poor cell state, etc., to increase the infection rate of cell lentivirus , Improve the culture and infection rate, reduce the effect of operation difficulty and cost

CN114561358APending Publication Date: 2022-05-31广州源井生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2022-05-31

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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.
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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...

Examples

Embodiment 1-5

[0072] A method for improving the infection rate of MDA-MB-231 cells, comprising the following steps;

[0073] (1) Digest the MDA-MB-231 cells in the logarithmic growth phase into a suspension single-cell liquid, and prepare 1×10 5 Amount of cell inoculation per well Inoculate the single cell suspension into a 12-well cell culture plate, add 1ml of conventional medium, and culture at 37°C and ambient air, wherein the conventional medium contains glutamine and Leibovitz's L-15 medium with fetal bovine serum.

[0074] (2) When the MDA-MB-231 cells in step (1) are cultured for 36 hours, replace the conventional culture medium with enhanced infection medium, and cultivate them at 37°C and in an environment with a volume fraction of 5% carbon dioxide for 24 hours , wherein the components of the enhanced infection medium are shown in Table 1 below;

[0075] (3) Maintain the culture conditions of step (2), use the lentivirus with EGFP expression box to infect MDA-MB-231 cells, the ...