Method for evoking differentiation of mesenchyma stem cell and fat stem cell into neurons and special culture medium thereof
A technology of adipose stem cells and mesenchymal stem cells, which is applied in the field of cell culture methods and its special medium, and can solve problems such as toxic and side effects
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Embodiment 1
[0037] Example 1. Inducing differentiation of mesenchymal stem cells into neurons
[0038] Taking mesenchymal stem cells derived from iliac bone marrow as an example, the differentiation induction effect of the method of the present invention was tested. Bone marrow mesenchymal stem cells can be obtained by the operation in the following step 1, or directly obtained from a stem cell bank.
[0039] 1. Isolation, purification, primary and subculture of bone marrow mesenchymal stem cells
[0040] Obtain iliac bone marrow from surgically discarded iliac bone marrow, or extract ilium bone marrow from healthy voluntary donors under sterile conditions, or obtain bone marrow from a bone marrow bank, add DMEM-F12 culture medium (high-glucose DMEM medium and DF-12 medium (product of Sigma Company, Cat#D0547) was obtained by mixing at a volume ratio of 1:1), centrifuged, discarded the supernatant and fat layer, and fully mixed the bone marrow cell pellet with DMEM-F12 medium Finally, g...
Embodiment 2
[0084] Example 2. Inducing the differentiation of adipose-derived stem cells into neurons
[0085] Taking adipose-derived stem cells as an example, the differentiation induction effect of the method of the present invention was tested. Adipose stem cells can be obtained by the operation in the following step 1, or directly obtained from a stem cell bank.
[0086] 1. Isolation, purification, primary and subculture of adipose-derived stem cells
[0087] 1) Use the fat discarded during liposuction or absorb fat from healthy volunteers by liposuction, rinse with PBS, add 4 times the volume of 0.1% collagenase, 1 times the volume of 2% BSA and double antibody, and then Put it into a 50mL centrifuge tube, digest at 37°C for 45min, and shake it from time to time.
[0088] 2) Filter the adipose tissue that has been digested into a paste with an 80-mesh sieve, and then filter it again with a 200-mesh sieve. Add collagenase to the remaining adipose tissue to continue digestion and rep...
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