In-vitro separation culture method for skeleton satellite cells of excellent ice and snow athletes

A technology of satellite cells and separation methods, applied in the interdisciplinary fields of sports traumatology, cell biology, and tissue engineering

Inactive Publication Date: 2013-06-19
HARBIN INST OF PHYSICAL EDUCATION
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  • Abstract
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Problems solved by technology

At the same time, traditional treatment methods, such as open surgery, often bring unpredictable risks for athletes to return to the peak level of competition due to side injuries and postoperative complications

Method used

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Examples

Experimental program
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Effect test

Embodiment Construction

[0010] (1) Take a 3mm biopsy tissue from the deltoid muscle of an excellent ice and snow athlete 3 ;

[0011] (2) Wash twice with 0.9% sterile saline;

[0012] (3) digest with 1% type I collagenase for 30min;

[0013] (4) digest with 0.1% pronase for 30 minutes, and separate the satellite cells from the basement membrane by pipetting with burettes of different specifications;

[0014] (5) Terminate the digestion with high-sugar DMEM medium containing 10% fetal bovine serum, filter with a 400-mesh cell sieve, and centrifuge at 500 g for 5 min.

[0015] (6) Discard the supernatant, resuspend in high-sugar DMEM medium, and inoculate for adherent culture.

[0016] (7) When the adherent and confluent rate of skeletal muscle satellite cells reached 95%, they were digested and subcultured with 0.25% trypsin.

[0017] (8) Cryopreservation of skeletal muscle satellite cells was performed using cryopreservation solution DMEM basal medium + dimethyl sulfoxide + fetal bovine serum (ra...

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Abstract

The invention relates to an in-vitro separation culture technique for the skeleton satellite cells of excellent ice and snow athletes, and belongs to the interdiscipline field of cytobiology, sports traumatology and tissue engineering. The technique comprises the following steps: cleaning muscular tissues by using 0.9% sterile saline solution; digesting and separating the skeleton satellite cells under the cooperative action of 1% of I-type collagenase and 0.1% of pronase under the condition of 37 degrees; culturing the skeleton satellite cells by using a high-glucose DMEM (Dulbeccos Modified Eagles Medium) culture medium containing 10% of fetal calf serum; and freezing and storing the skeleton satellite cells by using a freezing-stored liquid formed by the DMEM basal culture medium, dimethyl sulfoxide and the fetal calf serum which are in the ratio of 5:1:4. Thus, the simple, efficient and stably-inherited in-vitro separation cultivation method for the skeleton satellite cells of the excellent ice and snow athletes is provided.

Description

technical field [0001] The invention relates to an in vitro separation and culture technique for skeletal muscle satellite cells of excellent ice and snow athletes, and belongs to the interdisciplinary fields of sports traumatology, cell biology and tissue engineering. Background technique [0002] Skeletal muscle satellite cells (skeleton satellite cells, SCCs) are currently recognized skeletal muscle stem cells, which are myogenic cells with proliferation and differentiation potential located between the muscle cell membrane and the basement membrane. In 1961, Mauro described a cell closely connected to the edge of frog muscle fiber cells, and named it satellite cells according to their location. They are generally in a resting state, and when activated, they have the ability to proliferate, differentiate, and fuse into muscle cells. Ability to regenerate and regenerate muscle cells. Responsible for skeletal muscle growth and injury repair. In 2008, Nature magazine publi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
Inventor 王莹谢长福张宏宇熊慧孙婷婷关伟军
Owner HARBIN INST OF PHYSICAL EDUCATION
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