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Method for verifying spontaneous transformation of mice adipose-derived stem cells

An adipose stem cell and identification method technology is applied in the identification field of spontaneous transformation of mouse adipose stem cells, which can solve the problems of easy changes in biological characteristics and lack of detection indicators.

Inactive Publication Date: 2014-10-01
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] As an important model animal, mice are widely used in basic research. However, when adipose stem cells are cultured in vitro, their biological characteristics are easily changed. For how to effectively identify whether mouse adipose stem cells have undergone spontaneous transformation, There is still a lack of comprehensive and typical detection indicators

Method used

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Embodiment Construction

[0020] In order to make the technical problems to be solved and the beneficial effects of the present invention clearer, the present invention will be further described in detail below in conjunction with the examples.

[0021] One. First, the isolation and cultivation of mouse adipose stem cells comprises the following steps:

[0022] (1) Get 4-week-old ICR male mice, kill them, and disinfect their whole bodies;

[0023] (2) Cut out the adipose tissue at the groin under aseptic conditions, put it into DMEM / F12 to wash away the blood stains;

[0024] (3) Shred fully, add digestive juice, digest at 37°C for 1 hour, the composition of digestive juice is 10% fetal bovine serum (FBS), 0.09% collagenase Ⅰ, and the balance is DMEM / F12;

[0025] (4) After digestion, centrifuge, discard the upper layer of mature adipocytes and the middle layer of the digestive fluid, and then resuspend the bottom layer of cells with DMEM / F12;

[0026] (5) The cell suspension is successively filtered...

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Abstract

The invention discloses a method for verifying the spontaneous transformation of mice adipose-derived stem cells. The method comprises the step of in-vitro continuous passage of the mice adipose-derived stem cells and then comprises the following five steps: morphological change, sharp proliferation speed rise, abnormal chromosome generation and proportion increase, DNA (Deoxyribose Nucleic Acid) content reduction and surface molecule change. According to the method, the observation and detection of morphology and proliferation speed can be used for primarily judging whether the mice adipose-derived stem cells are transformed or not; the karyotype analysis of chromosomes can be used for analyzing whether the mice adipose-derived stem cell chromosomes are abnormal or not at a local level, the DAN content analysis can be used for detecting the quantitative change of a nuclear matter at an integral level, and the two methods most explain whether the mice adipose-derived stem cells are transformed or not; after the mice adipose is transformed, the abnormal expression of the representative molecules of the mice adipose-derived stem cells is caused, wherein the abnormal expression comprises the change of CD44, CD105 and Sca01; and conversely, the transformation of the mice adipose-derived stem cells can also be judged by monitoring the change of surface molecules in late-stage study, and the method is simpler, more direct and more effective.

Description

technical field [0001] The invention relates to a method for identifying spontaneous transformation of mouse adipose stem cells. Background technique [0002] Adipose stem cells have the potential of multidirectional differentiation, and can be directional differentiated into fat, osteoblast, cartilage, liver, cardiomyocytes, etc. in vitro. effect, so it is of great significance. [0003] As an important model animal, mice are widely used in basic research. However, when adipose stem cells are cultured in vitro, their biological characteristics are easily changed. For how to effectively identify whether mouse adipose stem cells have undergone spontaneous transformation, There is still a lack of comprehensive and typical detection indicators. Contents of the invention [0004] The technical problem to be solved by the present invention is to provide an identification method for the spontaneous transformation of mouse adipose stem cells, which provides a comprehensive, sim...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12Q1/68C12Q1/04G01N15/14
Inventor 张运海魏超李运生章孝荣方富贵张鹏飞李侠姜少帅韩菲宋丹丹刘通
Owner ANHUI AGRICULTURAL UNIVERSITY
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