Study on the preparation process of human umbilical cord mesenchymal stem cells
Human umbilical cord mesenchymal stem cells have shown varying degrees of efficacy in the treatment of a variety of refractory diseases,and the quality of stem cells is an important guarantee of efficacy.At present,different laboratories have different methods and key parameters for the preparation of stem cells,resulting in different quality of prepared stem cells.How to establish a set of scientific,stable and high-quality human umbilical cord mesenchymal stem cell preparation process is the goal of stem cell preparation engineering team.To establish a standardized preparation process for human umbilical cord mesenchymal stem cells(hUC-MSCs).Different culture media,digestion solutions,and digestion times were compared to evaluate the proliferation level,phenotype,and differentiation potential of hUC-MSCs.Different cell seeding densities and confluence rates were designed for cultivation,and the cell growth status and doubling time were observed.hUC-MSCs showed better proliferation level,phenotype,osteogenic,and adipogenic abilities when cultured in MSC-T4 medium with a 3-minute digestion using CTS enzyme.The optimal cell recovery seeding density and passage seeding density for hUC-MSCs were determined to be 10 000 cells/cm2 and 8 000 cells/cm2,respectively.A confluence rate of 90％～95％was found to be optimal for hUC-MSCs passage.
human umbilical cord mesenchymal stem cells(hUC-MSCs)preparation technologyculture mediadigestion solutionsseeding density
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