A preparation method and kit for clinical-grade human induced pluripotent stem cell-derived mesenchymal stem cells
A technology of pluripotent stem cells and mesenchymal stem cells, which is applied in the field of preparation of clinical-grade human induced pluripotent stem cell-derived mesenchymal stem cells, can solve the problems that mesenchymal stem cells cannot be used, and achieve the effect of unified source
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[0034] The invention provides a method for preparing clinical-grade human induced pluripotent stem cell-derived mesenchymal stem cells, comprising the following steps:
[0035]1) digesting human induced pluripotent stem cells to make a single cell suspension, and inoculating it on a pretreated culture plate;
[0036] 2) When the cells reach 80% confluence, use the induction medium for induction culture, and change the medium every day;
[0037] 3) When the cells reach 100% confluence, they are digested to make a single cell suspension, inoculated on the expansion medium for expansion culture, and the cells obtained after 3 to 4 generations of expansion culture are clinical-grade Mesenchymal stem cells.
[0038] In the present invention, human induced pluripotent stem cells are digested to make a single cell suspension, which is seeded on a pretreated culture plate.
[0039] In the present invention, the preparation method of human induced pluripotent stem cells is to separat...
Embodiment 1
[0049] The formula of MSC induction medium (referred to as M+S+C2):
[0050] 1L α-MEM basal medium contains 10% human platelet extract, 10 μmol / L SB431542 and 2 μmol / L CHIR99021. MSC induction medium was obtained after conventional sterilization.
Embodiment 2
[0052] Recipe of MSC Induction Medium:
[0053] 1L α-MEM basal medium contains 10% human platelet extract, 5 μmol / L SB431542 and 5 μmol / L CHIR99021. MSC induction medium was obtained after conventional sterilization.
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