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

Abstract

The invention discloses a culture method of tumor stem-cell-like cells of a rat pituitary prolactinoma cell strain. The culture method comprises the following steps: 1) taking cells in good condition in logarithmic growth phase; 2) inoculating the cells into a culture plate at a proportion of 100,000-300,000 cells / ml of culture medium; 3) carrying out intermittent culture for 1 day, then centrifuging for 5min at 1000rpm under 4 DEG C, discarding a half of supernatant liquid and adding the equal amount of a new stem cell culture medium; 4) culturing the stem cells for 2-3 days until cell debris appears in the culture medium, and further changing the half of the liquid every other day; 5) culturing for one week until small cell spheres in good condition appear; 6) culturing for one week until stem cell spheres are initially formed and cell proliferation is arrested; and 7) further culturing for two weeks until the stem cell spheres are basically formed. By separating the tumor stem-cell-like cells from the rat pituitary prolactinoma cell strain, the method disclosed by the invention can be used for solving the problem that the tumor stem-cell-like cells are difficult to obtain, and the method can be used for providing sufficient tumor stem-cell-like cells for further study.

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 adenomas are benign intracranial tumors, which can be classified according to endocrine hormone levels: prolactinoma, growth hormone adenoma (GH adenoma), corticotropin-releasing hormone adenoma (ACTH adenoma), thyroid-stimulating hormone gland adenoma (TSH adenoma), gonadotroph adenoma (LH / FSH adenoma) and nonfunctioning 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 ...

AI Technical Summary

Problems solved by technology

[0005] Pituitary prolactinomas account for about 30% of pituitary tumors, and most of them are treated with drugs (International Pituitary Society Prolactinoma Diagnosis and Treatment Guidelines, Guidelines of the Pituitary Society for the diagnosis and management of prolactinomas. Clin Endocrinol (Oxf), 2006, 65:265-273.), relatively few cases underwent surgery, and pituitary prolactinomas are benign tumors. The specimens obtained during the surgery were cultured in vitro, and the cells grew slowly, and generally stagnated after 3-4 passages. However, pituitary prolactinoma stem cell-like cells need to be separated and cultured from tumor cells, so there are problems of few specimen sources and difficult culture

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