Culture method of tumor stem cell-like cells of rat pituitary prolactinoma cell line

Abstract

The invention discloses a method for culturing tumor stem cell-like cells of a rat pituitary prolactinoma cell line: 1) taking cells in the logarithmic growth phase in a good state; 2) inoculating at a ratio of 10-300,000 cells / ml culture medium 3) After culturing for one day, centrifuge at 1000 rpm at 4°C for 5 minutes, discard half of the supernatant, and re-add the same amount of new stem cell culture medium; 4) After 2-3 days of stem cell culture, cell debris appears in the culture medium 5) After one week of culture, small cell spheres in good condition appeared; 6) After one week of culture, the stem cell spheres were initially formed and the cell proliferation stagnated; 7) After two weeks of culture, the stem cell spheres were basically formed. The invention separates tumor stem cell-like cells from rat pituitary prolactinoma cell lines, thereby solving the problem of few sources of tumor stem cell-like cells, and can provide a large amount for further research.

Description

technical field [0001] The invention relates to a method for culturing tumor stem cell-like cells, in particular to a method for cultivating tumor stem cell-like cells of a rat pituitary prolactinoma cell line. Background technique [0002] Pituitary adenoma is a benign intracranial tumor, which can be divided into prolactinoma, growth hormone adenoma (GH adenoma), corticotropin-releasing hormone adenoma (ACTH adenoma) and thyrotropin adenoma according to endocrine hormone levels. (TSH adenoma), gonadotroph adenoma (LH / FSH adenoma) and non-functioning adenoma (NFPA). [0003] The reports on tumor stem cells mainly focus on malignant tumors, and there are few reports on benign tumor stem-like cells. [0004] At present, there are 4 reports on the culture of pituitary adenoma stem cells in SCI indexed papers, 3 of which were isolated and cultured from human pituitary adenoma specimens, 2 of which were similar results published by the same research group, and the other was fro...

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Problems solved by technology

[0005] Pituitary prolactinomas account for about 30% of pituitary tumors, most of which are the first choice for drug treatment (Guidelines of the Pituitary Society for the diagnosis and management of prolactinomas. Clin Endocrinol (Oxf), 2006, 65 :265-273.), the cases of surgery are relatively few, and pituitary prolactinoma is a benign tumor. The specimens obtained during the operation are cultured in vitro, and the cells grow slowly, and generally after 3-4 generations, there will be growth stagnation, while Pituitary prolactinoma stem cell-like cells need to be separated and cultured from tumor cells, so there are problems of few samples and difficult culture

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