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目的 本研究拟从胎盘组织中分离培养获得间充质干细胞(placentaderivedmesenchymalstemcells,PMSCs) ,研究其生物学特性,探讨其在移植治疗方面的可行性。方法 将胎盘组织经胶原酶消化和贴壁培养获得的基质细胞,用免疫荧光标记和免疫细胞化学检测细胞表型,用活细胞记数和碘化丙啶(propidiumiodide,PI)检测其增殖能力,用叔丁对甲氧酚(butylatedhydroxyanisole ,BHA)、N2 培养基等对培养的细胞分别向神经和脂肪方向诱导分化,继而采用免疫细胞化学等方法对分化的细胞进行鉴定。结果 由胎盘组织中分离获得了一群具有与骨髓间充质干细胞(bonemarrowmesenchymalstemcells ,BMSCs)相似的形态和细胞表面标志的细胞:该群细胞增殖能力强;表达CD2 9、CD4 4、CD10 5 (SH2 )、ASMA、Ⅰ型和Ⅲ型胶原以及免疫应答负性调控分子PDL 1,但不表达CD11b、CD34、CD4 5、CD19、CD10 6 (VCAM 1)、CD117、HLA DR、vWF、MySM、HLA DR ,以及免疫应答正性调控分子CD4 0、CD4 0L、CD2 8、CD80和CD86 ;可诱导表达神经元标志神经微丝(neurofila ment,NF)和星型胶质细胞标志胶质纤维酸性蛋白(glialfibrilamentacidicprotein ,GFAP) ,并且这些细胞表达神经干细胞标志巢蛋白(nestin) ,向脂肪分化的细胞苏丹Ⅲ染色阳性。结论 本实验中获?
OBJECTIVE: To study the biological characteristics of placental derived mesenchymal stem cells (PMSCs) isolated from placenta tissue and investigate the feasibility of the transplanting treatment. Methods The stromal cells obtained by collagenase digestion and adherent culture were used to detect the cell phenotypes by immunofluorescence staining and immunocytochemistry. The proliferation of placental cells was detected by viable cell count and propidium iodide (PI) The cultured cells were induced to differentiate into neuronal and adipose tissue respectively by using butylated hydroxyanisole (BHA) and N2 medium, and then the differentiated cells were identified by immunocytochemistry and the like. Results A group of cells with similar morphology and cell surface markers to bone marrow-derived mesenchymal stem cells (BMSCs) were isolated from placenta: the cells proliferated well; the expression of CD2 9, CD4 4, CD10 5 (SH2) , ASMA, type I and type III collagen as well as immunoreactive negative regulatory molecule PDL 1 but not CD11b, CD34, CD4 5, CD19, CD106 (VCAM 1), CD117, HLA DR, vWF, MySM, HLA DR, As well as the positive regulatory molecules of immune response CD4 0, CD4 0L, CD2 8, CD80 and CD86; induced neurofilament (NF) and glial fibrillary acidic protein (glialfibrilamentacidicprotein, GFAP), and these cells expressed neural stem cell marker nestin and stained positive for Sudan III, a fat differentiated cell. Conclusion In this experiment,