Thalassemia-induced multipotent stem cell as well as preparation method and application thereof
A technology for pluripotent stem cells and thalassemia, which is applied in the field of thalassaemia-induced pluripotent stem cells and their preparation, and can solve problems to be improved, etc.
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[0051] 1. The acquisition and identification of original induced pluripotent stem cells
[0052] First, exosomes containing cDNA or miRNA clusters of pluripotent stem cell-inducing factors were introduced into the amniocytes of thalassemia patients by nucleofection method, wherein, compared with the normal β-globin gene, the thalassemia patients had The two alleles of the β globin gene in the amniocyte genome had the same mutation, the mutation type was IVS-II-654; the pluripotent stem cell induction factors used were Oct4, Sox2, Nanog, Lin28 and miR- A combination of 302-367.
[0053] Then, the amniotic fluid cells introduced with pluripotent stem cell-inducing factors were directly transferred into a serum-free, cell-free feeder layer human embryonic stem cell culture environment, and after 20 days of culture, a large number of typical primary induced pluripotent stem cell clones were obtained, and these clones were picked Then continue to cultivate. Two obtained original ...
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