人胎盘血间质干细胞分离培养

目的：分离培养人胎盘血间质干细胞（hMSC），为hMSC探寻新来源。方法：采用羟乙基淀粉（HES）方法分离、富集胎盘血有核细胞；DMEM培养液体外培养、纯化、扩增hMSC；流式细胞仪检测细胞表面标记；地塞米松、IBMX、胰岛素和吲哚美辛定向诱导hMSC样细胞向脂肪细胞分化。结果：胎盘血来源有核细胞,在DMEM体外培养条件下，生长出具有塑料粘附特性的梭形细胞，阳性获得率29.17%（7/24），该细胞传代培养达6个月以上；流式细胞仪检测结果显示CD29、CD44、CD59、CD90、CD105、CD166表达阳性，CD14、CD34、CD45、CD80、CD86表达阴性；加入脂肪细胞诱导剂，细胞在形态上向脂肪细胞转化，胞内出现脂滴、脂泡，油红O阳性。结论：人胎盘血可以分离培养出hMSC，是hMSC的重要来源。

Isolation of mesenchymal stem cells from human umbilical cord blood

AIM: To investigate the biological characterics of human second-trimester fetal cord blood mesenchymal stem cells (MSC) and its application prospects in utero gene transfer/therapy (IUGT). METHODS: Nuclear cells separated from cord blood were cultured in DMEM medium. Surface antigens of the MSC were analyzed by the FACScan flow cytometry. Adipogenic and osteogenic mediums were used to assess the differentiation ability of the cells. Adenovirus vector deliver green fluorescent protein gene (Ad-GFP) was used to transfected the MSC and the expressing of GFP was detected by fluorescent microscope. The MSC were injected into the liver of newborn rat. The immunofluorescence analysis was conducted to determine the presence of double-positive CD105⁺/CD166⁺ cells in different organs of rats. MSC were subcutaneous injected into the human-nonobese diabetes/severe combined immunodeficiency disease (NOD/SCID) mice and carcinogenesises of the MSC in vivo were detected by pathological diagnosis. RESULTS: MSC could be separated from fetal cord blood. These cells were uniformly positive for CD29, CD44, CD59, CD105, CD166 and negative for CD34, CD45, CD80, CD86, HLA-DR. The cells had the abilities to differentiate into adipogenic and osteogenic cells in vitro, expressed the GFP at high levels (56.32%±3.28%). The MSC were located at different organs after injected into the newborn rats and didn't have carcinogenicity in vivo. CONCLUSION: Human second-trimester fetal cord blood MSC is an promising target cells in fetal IUGT.