Comparison Of Biological Characteristics Of MSCs Derived From Human Adipose And Lipoma

Objective: 1.To extract and isolate the mesenchymal stem cells derived from human adipose tissue and lipomas by enzyme digestion method,and identify and culture the stem cells in vitro.2.To compare the biological characteristics of mesenchymal stem cells derived from human adipose tissue and lipoma tissue,it is helpful to a further study on human lipoma-derived mesenchymal stem cells(hLMSCs),and to provide the basic data for exploring the occurrence mechanism and identification of lipoma.Methods:Human adipose-derived mesenchymal stem cells(h ASCs)and hLMSCs were isolated from the same volume of fat and lipoma which was digested by collagenase and cultured.Morphology and growth condition of h ASCs and hLMSCs were observed with electron microscopy.To further demonstrate the power of adipogenic plasticity of hASCs and hLMSCs,h ASCs and hLMSCs were promoted osteogenic differentiation and adipogenic differentiation.The cell surface marker CD34,CD45,CD90 and CD133 were detected by flow cytometry.Proliferation of hASCs and hLMSCs were examined by CCK8 technique.Expression of HMGA2 in hASCs and hLMSCs by quantitative real-time PCR..Results:1.The mesenchymal stem cells derived from fat and lipoma were isolated and purified by collagenase.It was found that:(1)hASCs and h LMSCs finished adhesive growth after24 hours.(2)The first fusion as conglobation: Primary cells were extracted and it was found that the number of the stem cells of lipoma tissue more than the number of stem cells extracted from adipose tissue,but the time of first fusion as conglobation of hASCs and hLMSCs was basically the same.The cells were reached growth peak at 6~8 days in primary culture(3)The hASCs and hLMSCs of the primary culture had a large number of heterozygous cells,and the cells were gradually disappearing after cell fusion.Both cells grew spindle-shaped and appeared in a spiral-like arrangement.(4)Two cells in osteogenic adipogenic induction can be differentiated into adipocytes and bone cells.(5)The expression of CD45 and CD90 on the surface markers of hASCs and h LMSCs were detected by flow cytometry.The results showed that hASCs and hLMSCs had the same characteristics.The expression of CD34 and CD133 in hASCs were 2.14% and 3.17%,respectively.The expression of CD34 and CD133 in hLMSCs were 20.62% and 16.04%,respectively.(6)The CCK8 assay's results unveiled that the hLMSCs were capable of a faster growth and proliferation compared to hASCs.At the fourth-day time-points,a noteworthy dissimilarity was witnessed between hASCs and h LMSCs,and the cell proliferation of hLMSCs was higher than hASCs.(7)The expression of HMGA2 in hASCs and hLMSCs group were detected by real-time quantitative PCR.The expression of HMGA2 in h LMSCs group was significantly higher than that in hASCs group.Conclusions: The hASCs and hLMSCs were isolated and cultured with collagenase digestion.Both hASCs and h LMSCs were adherent and grown.After induction,they had osteogenic and adipogenic ability,which had the potential of multi-directional differentiation and expressed similar cell surface markers.And the expression of HMGA2 were different,which show significant understanding for the development of lipoma and the development of benign tumor.