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

A prolactinoma, tumor stem cell technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problems of few specimen sources, slow cell growth, and difficulty in culturing

Active Publication Date: 2015-03-25
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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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 s...

Method used

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

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Embodiment 1

[0025] 1. Stem cell culture medium formula:

[0026]

[0027]

[0028] 2. Culture conditions: 5% CO2, 37°C;

[0029] 3. Training methods and key points:

[0030] 1) Take cells in the logarithmic growth phase in good condition (Figure A), wash 2-3 times with PBS, and resuspend with the above-mentioned stem cell medium;

[0031] 2) Inoculate in the culture plate at a ratio of 100,000-300,000 cells / ml culture medium. It is recommended to use a 24-well plate for culture, which has the highest efficiency, followed by a 6-well plate;

[0032] 3) After interval culture for one day, centrifuge at 1000rpm at 4°C for 5min, discard half of the supernatant, and re-add the same amount of new stem cell culture medium;

[0033] 4) After the stem cells were cultured for 2-3 days, a large number of cells were apoptotic (Figure B), and cell fragments appeared in the medium, so continue to change the medium every other day;

[0034] 5) After 1 week, small cell spheres in good condition ...

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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 ...

Claims

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Application Information

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IPC IPC(8): C12N5/095
Inventor 苏志鹏蔡霖陈贤斌陈健卢江龙王成德李群
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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