胎盘、脐血及骨髓来源贴壁细胞生物学特性比较.docVIP

胎盘、脐血及骨髓来源贴壁细胞生物学特性比较 　【摘要】 　　目的 探讨人胎盘、脐血和骨髓来源的贴壁细胞的分离培养和生物学特性，为间充质干细胞(MSC)的选择和应用提供依据。方法 采用酶消化法分离人胎盘组织，60g/L羟乙基淀粉和密度梯度离心两步法分离脐血单个核细胞，密度梯度离心法分离骨髓单个核细胞，分别进行贴壁培养，观察不同来源MSC的生长、增殖和表面标志的表达并做比较。结果 胎盘来源的贴壁细胞为一种混合性细胞，脐血单个核细胞体外培养可以获得形态不均一的贴壁细胞，二者在生长规律和形态学特征上与骨髓基质相比有一定差异性，CD106、CD44在三种细胞均有表达。结论 胎盘和脐血来源的贴壁细胞具备间充质干细胞的基本特征。 【关键词】 胎盘；脐血；间充质干细胞 　　Comparison of the adherent cells derived from human placenta,umbilical cord blood and bone marrow 　　ABSTRACT: Objective To compare the adherent cells derived from human placenta, umbilical cord blood and bone marrow, and provide laboratory data for clinical application of mesenchymal stem cells. Methods Adherent cells were isolated from human placenta tissues by enzyme digestion, and mononuclear cells (MNC) were isolated from umbilical cord blood (UCB) by 60g/L HES and density gradient centrifugation and MNC were isolated from bone marrow (BM) by density gradient centrifugation, and then these cells were cultured in vitro. Their biological characteristics were studied and compared. Results The adherent cells cultured from human placenta and umbilical cord blood had disparate shape in vitro respectively. And they had some differences in growth and shape from those derived from bone marrow. The adherent cells derived from the three tissues all expressed CD106 and CD44 in immunohistochemistry staining. Conclusion The adherent cells derived from human placenta and umbilical cord blood have the basic features of mesenchymal stem cells. 　　KEY WORDS: placenta; umbilical cord blood; mesenchymal stem cell 　　间充质干细胞(mesenchymal stem cell, MSC)是成体干细胞之一，不仅支持造血系统，还具有多向分化潜能，在一定的实验条件下可分化为成骨细胞、软骨细胞、脂肪细胞、神经细胞、肌细胞、心肌细胞、肝细胞等[15]。因此，在细胞微环境和组织工程等方面具有广泛的应用前景。近年来，已有具有干/祖细胞特征的间充质细胞自骨髓、外周血、软骨、肌肉等组织中分离出来。这些组织均来源于胚胎发育期的中胚层，胎盘由滋养细胞及大量起源于中胚层的间充质和血管共同组成，亦有报道从脐带中分离培养出间充质干细胞[67]。因此，提示我们，与脐带同为胎儿附属物的胎盘中可能也含有MSC。本实验试从胎盘和脐血中培养出类似于骨髓来源的间充质干细胞的贴壁细胞，并对这三种细胞进行形态学观察，利用免疫细胞化学方法对其进行表面抗原的测定和比较，探讨其作为MSC新来源的可行性。 　　1 材料与方法 　　1.1 材料 胎盘、脐血均采自本院产房。供者要求是身体健康的产妇，胎儿发育良好。新鲜骨髓标本采自本院门诊营养性贫血(骨