RESUMO
BACKGROUND:Culture condition,isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells,which lack of unified identification standards.Therefore,it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem calls(hUCMSCs).OBJECTIVE:To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts.METHODS:The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase.The morphology,proliferation and immunophenotype of the 3rd passage cells were analyzed,and then cells were induced to osteogenic and adipogenic differentiation in vitro.RESULTS AND CONCLUSION:The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro.Flow cytometry analysis revealed that the hUCMSCs were positive for CD29 CD44,CD59,CD105,but were negative for CD40,CD86 and HLA-DR.These calls could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions.The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro.This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts.
RESUMO
Purpose The aim is to invest the mechanism and activity of kringle1～3 gene of angiostatin. Methods Kringle1～3 gene of angiostatin was abtained from hepatocyte of normal human by RT-PCR and inserted into expression vector pPIC9K in Pichia expression system. The recombinant protein was induced secrete , purified and test the activity by MTT.Results Krigle1～3 protein could inhibit vascular endothelial cell proliferative activity.Conclusion The reaction similar apoptosis induced by kringle1～3 recombinant protein of angiostatin inhibit cell proliferation.