Method for separation and purification, long-term passage cultivation, cryopreservation and recovery of sheep spermatogonia stem cells
A technology for the separation and purification of spermatogonial stem cells, which is applied in the fields of separation and purification, cryopreservation and recovery, passage and long-term culture of sheep spermatogonial stem cells, and can solve the problems such as difficult separation and purification of spermatogonial stem cells
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[0081] The technical solution of the present invention is further described below through specific examples, but the technical solution of the present invention is not limited to the examples.
[0082] A method for separating and purifying sheep spermatogonia stem cells of the present invention comprises the following steps:
[0083] Step 1: Prepare the solution:
[0084] 1) Add 0.1% antibacterial-antifungal agent by volume to DMEM / F12 culture solution to make culture solution A, and use it within two weeks;
[0085] 2) Add type IV collagenase at a concentration of 20mg / ml to DMEM low-sugar culture medium to prepare type IV collagenase stock solution, and store at -20°C;
[0086] 3) Add hyaluronidase at a concentration of 10 mg / ml to DMEM low-sugar culture medium to prepare hyaluronidase stock solution, and store at -20°C;
[0087] 4) Add DnaseI to DMEM low-sugar culture medium at a concentration of 5 mg / ml, and then add glycerol with a volume percentage of 20% to prepare a ...
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