【摘要】 从胎盘中分离培养人胎盘源贴壁细胞 (humanplacentaderivedadherentcells,hPDAC) ,研究其对脐血CD34+细胞体外扩增作用。以酶消化法自人胎盘组织中分离培养hPDAC ,并以流式细胞术对其进行鉴定。进一步 ,采用免疫磁珠法分离人脐血CD34+细胞 ,建立以hPDAC为滋养层的CD34+细胞体外扩增培养体系 ,并与无滋养层的液体培养体系相比 ,观察不同培养体系对有核细胞总数、CFC及CD34+细胞百分率的影响。结果表明 ,人胎盘组织中可分离培养出hPDAC ,并证实其为混合性细胞群体 ,主要含有间充质细胞。以hPDAC为滋养层的共培养体系较无滋养层液体培养体系对有核细胞总数、CFC及CD34+细胞均具有明显的扩增效应 ,其中以SCF +IL 3+IL 6 +FL +hPDAC组扩增效果最强 ,分别扩增了 ( 12 6 .0± 6 .7)倍 ,( 4 9.8± 1.7)倍和 ( 8.3± 1.6 5 )倍。上述结果提示 ,hPDAC具有体外支持造血作用 ,并提供了一与脐血CD34+细胞体外扩增相适应的新滋养层。更多还原

【Abstract】 The objective of this study was to elucidate the effect of human placenta adherent cells (hPDAC) on expansion of human umbilical cord blood CD34 + cells in vitro. hPDAC was isolated and characterized in human placenta tissue by using enzyme digesting method and flow cytometry. A co culture system was established with hPDAC and cord blood CD34 + cells. The CD34 + cells were cultured in different culture systems with different combinations of hPDAC and SCF, IL 3, IL 6 and FL. The number of total nucleated cells, CFC and CD34 + cells were repeatedly counted in culture for 4 weeks. The results showed that the hPDAC displayed fibroblast like morphology, and were positive for CD29, CD44, CD166, HLA ABC and UEA 1, and negative for CD34, CD45 and HLA DR. Functionally, ex vivo expansion of CD34 + cells on feeder layer of placental adherent cells was significantly higher than that no feeder layer group. SCF+IL 3+IL 6+FL+hPDAC manifested the most potent combination, with the number of total nucleated cells increasing by (126.0±6.7) fold, progenitor cells (CFC) (49.8±1.7) fold and CD34 + cells (8.3±1.65) fold. It is concluded that placental adherent cells could support hematopoiesis in vitro and work as a suitable feeder layer for cord blood stem/progenitor cell expansion.更多还原