人胎盘绒毛膜来源间充质干细胞的生物学特性

背景：骨髓是间充质干细胞的主要来源,但骨髓中间充质干细胞数量和质量会随着年龄的增长而逐渐降低。因此,寻找一种新的干细胞来源具有重要意义。目的：分离研究了一种新的间充质干细胞来源——人绒毛膜来源间充质干细胞,并研究其生物学特性。方法：将胎盘冲洗干净后,分离出脐带和羊膜组织,将剩余的胎盘组织进行原代和传代培养。通过短串联重复序列分析检测细胞是否来源于绒毛膜,用MTT法检测细胞生长方式,流式细胞仪分析细胞表型及干细胞标记,使用不同的诱导分化培养基检测其多向分化的能力。将该细胞与植物血凝素刺激的外周血单个核细胞共培养,用酶联免疫吸附试验检测上清γ-干扰素的水平。结果与结论：短串联重复序列分析证明所得到的细胞来源于绒毛膜,这种绒毛膜来源的细胞生长呈典型的成纤维细胞形态。细胞表达常见的间充质干细胞表面标记CD90、CD73、CD105、CD44,不表达CD45,CD11b和CD34。同时,细胞也表达Nestin和Sox-2。在不同的条件培养基培养状态下,细胞可向成骨、成脂方向分化。这些结果证明所得到的细胞为人绒毛膜间充质干细胞。这些绒毛膜间充质干细胞能抑制植物血凝素刺激的人外周血单个核细胞分泌γ-干扰素。可见绒毛膜来源的间充质干细胞具有和传统的骨髓来源间充质干细胞具有相似的生物学特性。

Biological characteristics of human chorionic-derived mesenchymal stem cells

BACKGROUND：Bone marrow is major source of mesenchymal stem cells（MSCs）,but the quantity and quality of MSCs in bone marrow have been shown to be gradually reduced with the increase of age.Therefore,it＇s very important to find a new source of stem cells.OBJECTIVE：To study a new source of MSCs isolated from human chorionic villous and to study its biological characteristics.METHODS：Umbilical cord and amniotic membrane were isolated from clean placenta;the remaining placenta tissue underwent primary culture and subculture.Tandem repetitive sequence（TRS） analysis was used to detect whether the MSCs obtained from placenta is chorionic villous original.Cell growth pattern was detected by MTT assay.Flow cytometry was applied to analyze cell phenotype and stem cell markers.The different differential medium was used to detect its multi-directional differentiation ability.The immunomodulatory properties of these chorionic-derived cells were evaluated by coculturing the cells with peripheral blood mononuclear cells stimulated by phytohemagglutinin（PHA）,and the level of interferon-γ（IFN-γ） was detected by enzyme-labeled immunosorbent assay（ELISA）.RESULTS AND CONCLUSION：TRS analysis shows that these chorionic-derived cells are derived from human chorionic.They are adherent cells with typical fibroblast morphology.Flow cytometric analysis reveals that human chorionic-derived cells are consistently positive for MSCs labeled with CD73,CD29,CD44,CD90 and CD105,and negative for the hematopoietic markers CD45,CD11b and CD34.The cells can differentiate into osteoblasts and adipogenesis under different conditioned medium cultured conditions.These results proved that these cells are human chorionic-derived MSCs.The human chorionic-derived MSCs can inhibit peripheral blood mononuclear cell stimulated by PHA to secrete γ-interferon.