Preparation method of tumor suppressor gene-modified mesenchymal stem cells
A mesenchymal stem cell and gene modification technology, which is applied in the field of mesenchymal stem cell preparation, can solve the problems of immunogenicity limiting repeated administration, low immunogenicity, limiting the development and application of tumor gene therapy, and solving the problem of inability to efficiently target Delivery and safety issues, high infection rate and co-expression efficiency, good tumor targeting effects
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[0064] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.
[0065] Isolation and culture of mesenchymal stem cells (MSCs)
[0066] (MSCs include but are not limited to MSCs derived from human fat, umbilical cord, cord blood, placenta, bone marrow, dental pulp, etc. The example takes human adipose-derived mesenchymal stem cells-hAMSCs as an example)
[0067] (1) Isolation of hAMSCs and culture of primary and passage
[0068] Fat samples were obtained from healthy donors who underwent liposuction in cosmetic plastic surgery clinics. Take 20ml of adipose tissue obtained by liposuction, first digest it with 0.075% collagenase for 30 minutes, and filter the digested mixed liquid through a 200-mesh cell sieve, and wash the filtered cells twice with DMEM / F12 medium, respectively in Centrifuge at 1500rpm for 10 minutes at room temperature for cell enrichment, resuspend the cells in complete medi...
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