Primary mammary epithelial cell culture medium, culture method and application

Abstract

The invention provides a primary cell culture medium for culturing primary mammary epithelial cells and containing amphiregulin, and a culture method using the primary cell culture medium. Through the culture method, the primary cell culture medium is used for culturing primary cells on a culture vessel coated with extracellular matrigel, the primary cells grow on the culture vessel coated with the extracellular matrigel, and the primary cells are rapidly proliferated under the combined action of nutritional factors contained in the primary cell culture medium and extracellular matrigel. A cell model obtained by the primary cell culture medium and the primary cell culture method can be used for curative effect evaluating and screening of medicines.

Description

technical field [0001] The invention belongs to the technical field of medicine, in particular, it relates to a culture medium and a culture method for culturing or expanding primary mammary gland epithelial cells in vitro, and also relates to a method and application of the cultured cells in evaluating and screening the curative effect of drugs . Background technique [0002] Breast cancer is one of the most important malignant tumors affecting women's health. The latest statistical results show that globally, the incidence and mortality of breast cancer rank first and second in the incidence and mortality of female malignant tumors, respectively. In recent years, although people have made a lot of progress in the research on the molecular typing and pathogenesis of breast cancer, the current standard drug treatment methods for breast cancer are still based on hormones and cytotoxic drugs, lacking personalized precise drug guidance . Because of the diversity and complexi...

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Problems solved by technology

[0004] However, both techniques have certain limitations

Cell reprogramming is a technique in which the patient's own primary epithelial cells are co-cultured with murine feeder cells, however, the presence of these murine cells interferes with the patient's primary cells when performing drug susceptibility testing The drug sensitivity test results of autologous primary cells; however, if the mouse-derived feeder cells are withdrawn, the patient's autologous primary cells will be out of the reprogramming environment, and the cell proliferation rate and intracellular signaling pathways will be significantly changed (Liu et al., Am J Pathol, 183(6):1862-1870, 2013; Liu et al., Cell Death Dis., 9(7):750, 2018), so that the response of the patient's own primary cells to the drug is greatly affected

Organoid technology is a technology in which the patient's own primary epithelial cells are embedded in the extracellular matrix for three-dimensional culture in vitro. This technology does not require feeder cells, so there is no interference with mouse-derived feeder cells. However, the culture of organoid technology A variety of specific growth factors need to be added to the base, which is expensive and not suitable for popularization in clinics for large-scale application

In addition, during the whole culture process of organoids, cells need to be embedded in extracellular matrix gel, and the plating steps of cell seeding, passage and drug sensitivity testing are cumbersome and time-consuming compared with 2D culture operations, and the technology formed The size of organoids is not easy to control, and some organoids tend to grow too large and cause internal necrosis. Therefore, compared with 2D culture technology, organoid technology is less operable and applicable, and requires professional and technical personnel to operate. It is not suitable for large-scale and extensive clinical in vitro drug sensitivity testing (Nick Barker, Nat Cell Biol, 18(3):246-54, 2016)

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