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人胎盘作为间充质干细胞的新来源在调控造血干/祖细胞(CD34+细胞)发育与分化方面具有重要作用,但其在移植中是否对CD34+细胞迁移起作用尚不清楚.为此,本研究通过胰酶消化法和密度梯度离心法,单克隆筛选培养出人胎盘来源间充质干细胞,RT-PCR检测其与造血和趋化相关的细胞因子和黏附分子的表达;进一步利用transwell体系检测其对脐血CD34+细胞的迁移作用.结果表明,PDMSCs呈现典型的成纤维样细胞,表达间充质干细胞相关表面抗原CD73,CD90和CD105,不表达造血及内皮细胞表面抗原CD34,CD45,CD31等,PDMSCs具有诱导成脂肪、成骨和成软骨的能力.RT-PCR检测结果显示,PDMSCs表达IL-6,LIF,G-CSF,GM-CSF,M-CSF,FL,SCF,SDF-1,VEGF等多种造血与趋化因子,Integrinβ1,Integrinα5,ICAM-1,ICAM-2,ICAM-3,VCAM-1等多种黏附分子;将PDMSCs按照不同密度接种在transwell体系,4h后脐血CD34+细胞迁移率随着PDMSCs的密度增加而增加;在PDMSCs与SDF-1对CD34+细胞的趋化迁移比较实验中,PDMSCs对CD34+细胞的迁移作用明显高于单纯的SDF-1.上述结果提示,PDMSCs对脐血CD34+细胞具有明显的迁移作用,为今后探讨PDMSCs与脐血CD34+细胞共移植是否促进CD34+细胞的植入提供理论基础与实验依据.
As a new source of mesenchymal stem cells, human placenta plays an important role in regulating the development and differentiation of hematopoietic stem / progenitor cells (CD34 + cells), but it remains unclear whether it plays a role in the migration of CD34 + cells.In this study, The human placenta-derived mesenchymal stem cells were cultured by trypsin digestion and density gradient centrifugation. The expression of cytokines and adhesion molecules related to hematopoietic and chemotaxis were detected by RT-PCR. The transwell system was used to detect On the migration of cord blood CD34 + cells.The results showed that PDMSCs showed typical fibroblast-like cells, expressed CD73, CD90 and CD105 related surface antigen of mesenchymal stem cells, did not express hematopoietic and endothelial cell surface antigen CD34, CD45, CD31, The results of RT-PCR showed that PDMSCs could express IL-6, LIF, G-CSF, GM-CSF, M-CSF, FL, SCF, SDF- Such as integrinβ1, Integrinα5, ICAM-1, ICAM-2, ICAM-3 and VCAM-1. The PDMSCs were inoculated into transwell system at different densities. After 4h, CD34 + cells Mobility increases with increasing density of PDMSCs ; PDMSCs migration of CD34 + cells was significantly higher than that of pure SDF-1 cells in PDMSCs and SDF-1 compared to CD34 + cells. These results suggest that PDMSCs have a significant migration effect on cord blood CD34 + cells, To provide a theoretical basis and experimental evidence for exploring whether PDMSCs co-transplantation with cord blood CD34 + cells can promote the implantation of CD34 + cells in the future.