Methods for in vitro isolation and culture and induced differentiation of bovine muscle satellite cells
A muscle satellite cell and differentiation-inducing technology, which is applied in the field of in vitro isolation and differentiation of cells, can solve the problems of no contractile function, poor passage stability, and small number of myotubes, so as to save culture costs and improve differentiation For the effect of strong myotube ability and proliferative activity
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specific Embodiment approach 1
[0012] Specific embodiment one: the in vitro separation and culture method of bovine muscle satellite cells of the present embodiment is realized by the following steps: one, clean bovine muscle tissue with D-Hanks liquid and cut into 1mm 3 The small pieces were placed in D-Hanks solution containing 0.25g trypsin, digested at 37°C for 30min, then centrifuged to discard the supernatant, then added collagenase solution, digested at 37°C for 2-3h to obtain cells Suspension; 2. Use a 400-mesh sieve to filter the cell suspension, then centrifuge, and resuspend the resulting cell pellet with D-Hanks solution and centrifuge, then resuspend the cells with cell growth medium, and then inoculate in Polymeric Lyme In the cell culture flask coated with amino acid, the cells ppl to pp6 were cultivated by differential attachment method; 3. The cells in pp6 were cultivated to a confluence rate of 95%, and then passaged with trypsin with a mass concentration of 0.25%. Promptly complete the in...
specific Embodiment approach 2
[0026] Embodiment 2: This embodiment differs from Embodiment 1 in that the centrifugation in steps 1 and 2 is performed at a speed of 1000 r / min for 5 minutes. Other steps and parameters are the same as those in Embodiment 1.
specific Embodiment approach 3
[0027] Embodiment 3: The difference between this embodiment and Embodiment 1 is that in step 1, collagenase solution is added and digested at 37° C. for 2 hours to obtain a cell suspension. Other steps and parameters are the same as those in Embodiment 1.
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