Recoverable immortalized rat bone marrow mesenchyme stem cell as well as preparation method and application thereof
A bone marrow mesenchymal and stem cell technology, applied in the field of cell engineering, can solve the problems of lack of dynamic balance, easy degeneration, and short cell survival time, and achieve strong in vitro proliferation ability, stable biological properties, and good biological safety Effect
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[0030] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings.
[0031] The SSR69 plasmid and pAmhpo plasmid used in the preferred embodiment were donated by Professor He Tongchuan from the University of Chicago.
[0032] 1. Isolation, culture and identification of primary rat MSCs
[0033] SPF grade 4-8 week-old SD rats weighing 80-150 g (purchased from the Animal Center of Chongqing Medical University) were sacrificed by cervical dislocation. The femur and tibia were taken out under sterile conditions, and the bone marrow was washed out with DMEM / F12 medium, and centrifuged. Remove the supernatant and fat, press 3×10 per bottle 6 25cm 2 In a plastic culture bottle, use DMEM / F12 complete medium containing 10% fetal bovine serum at a temperature of 37°C and CO 2 Cultivate under the condition of 5% volume fraction, discard...
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