人脐带、胎盘绒毛膜来源间充质干细胞的生物学特性比较研究

Comparative Study of Biological Characteristics of Human Umbilical Cord and Placental Chorionic Villous Mesenchymal Stem Cells

由于组织来源和增殖能力的优势,与骨髓间充质干细胞(MSC)相比,脐带和胎盘来源的MSC更具有临床应用前景,但脐带和胎盘来源MSC的生物学特性是否有差异值得进一步研究。本研究比较人脐带、胎盘绒毛膜来源MSC的生物学特性。将胎盘、脐带冲洗干净后,通过酶消化法分离脐带、胎盘来源的MSC。通过短串联重复序列分析(STR)检测细胞是否均来源于胎儿组织,用MTT法检测细胞增殖能力,流式细胞术分析细胞表型,使用不同的诱导分化培养液检测其多向分化的能力。将MSC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。结果表明,STR分析证实所得到的细胞均来源于胎儿组织,这两种细胞生长呈典型的成纤维细胞形态;细胞表达常见的MSC表面标记CD90、CD73、CD105、CD44,不表达CD45,CD11b和CD34;在不同的条件培养液培养下,细胞均可向成骨、成脂方向分化,但在成脂分化方面,绒毛膜来源MSC能形成更大的脂滴。结论:所得到的细胞均为MSC,均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,且绒毛膜来源的MSC具有更强的抑制能力。这使得绒毛膜来源的MSC在治疗自身免疫系统疾病方面可能具有更大的优势。

Because advantage of tissue origin and proliferation potential,the umbilical cord-derived mesenchymal stem cells（UC-MSC） and placental chorionic villous-derived mesenchymal stem cells（CV-MSC） have clinical application potential,as compared with bone marrow MSC.But whether the differences of biological characteristics exist between UC-MSC and CV-MSC,which deserve to be further explored.This study was purposed to compare the biological chracteristics of UC-MSC and CV-MSC.The placental and umbilical cord were cleaned by using the sterile physiological salt,the UC-MSC and CV-MSC were separated by enzyme digestion.Short tandem repeat（STR） analysis was used to detect whether the MSC obtained from fetal tissue.MTT methed was used to detect proliferation of MSC.Flow cytometry was applied to analyze cell phenotype.The different differential medium was used to detect their multi-directional differentiation capacity.After the MSC and PHA-stimulated peripheral blood mononuclear cells were co-cultured,the γ-interferon（IFN-γ） levels of the co-culture supernatant were detected using the ELISA.The results showed that these MSC were derived from fetal tissue by STR amalysis.They were adherent cells with typical fibroblast morphology.Cells expressed the MSC surface markers CD90,CD73 and CD105 and CD44,not expressed CD45 and of CD11b and CD34.These cells could differentiate into osteoblasts and adipoblasts under culture with different conditioned medium,but in the adipogenic differentiation of CV-MSC,the larger lipid droplets appeared.It is concluded that these cells are obtained MSC.These MSC can inhibit peripheral blood mononuclear cells stimulated by PHA to secrete IFN-γ,and the the CV-MSC have a stronger suppression capacity,which makes the CV-MSC to have a greater advantage in the treatment of autoimmune diseases.