摘要
目的 :培养、鉴定、描述及扩增人脐血PSC344 5多潜能干细胞 .方法 :使用Ficoll Hypaque分离新鲜人脐血单个核细胞 ,用PBS洗涤 2～ 3遍后在Potentculture完全培养液中37℃ ,5 0mL/LCO2 条件下培养 1 0～ 1 4d ,得到一群成纤维样细胞 ,该细胞与荧光素标记的mAb (CD1 4 PE ,CD1 1b PE ,CD4 5 FITC ,CD34 PE ,CD3 FITC ,CD1 0 5 PE ,CD1 6 6 PE ,CD2 9 PE ,CD90 FITC ,CD33 PE ,CD4 1 PE ,CD4 4 FITC ,CD7 FITC ,CD6 1 FITC ,HLA DR PE)反应后 ,经流式细胞仪检测其表面标志 .并对该细胞进行了糖原 (PAS) ,酸性磷酸酶 (AP) ,碱性磷酸酶(Alk .P) ,油红O(ORO) ,过氧化物酶 (POX)等化学染色 .结果 :PSC344 5表达 :HLA DR+ ,CD1 4 + ,CD4 5 + ,CD1 1b+ ,CD2 9+ ,CD34-,CD1 0 5 -,CD3-,CD90 -,CD1 6 6 -.细胞化学染色为PAS+ ,AP+ ,Alk .P-,ORO-,POX-.在适当诱导剂的诱导下可以定向分化 .结论
AIM: To culture, identify, characterize and propagate human umbilical cord blood PSC3445 pluripotent stem cells. METHODS: Fresh human umbilical cord blood mononuclear cells were obtained after Ficoll Hypaque fractionation and two to three washes with PBS and the cells were incubated at Potentculture for 10-14 d at 37℃ (50 mL/L CO 2). The cells were determined by FACScan flow analysis with CD14 PE, CD11b PE, CD45 FITC, CD34 PE, CD3 FITC, CD105 PE, CD166 PE, CD29 PE, CD90 FITC, CD33 PE, CD41 PE, CD 44FITC, CD 7FITC, CD61 FITC and HLA DR PE, and stained for PAS, AP, Alk.P, and POX. RERULTS: PSC3445 cells expressed CD14, CD45, CD29, HLA DR and CD11b, but did not express CD34, CD105, CD166, CD90 and CD3. The cells displayed PAS +, AP +, Alk.P -, ORO - and POX - by special chemical staining and could be induced to differentiate into T cells, macrophages, epithelial cells, neuronal like cells by appropriate cell growth factor. CONCLUSION: The cells, fibroblasts like cells, are novel pluripotent stem cells in human umbilical cord blood.
出处
《第四军医大学学报》
北大核心
2004年第11期964-968,共5页
Journal of the Fourth Military Medical University
关键词
脐血
多潜能干细胞
单个核细胞
细胞分化
umbilical cord blood
Pluripotent stem cells
mononuclear cells
cell differentiation
