兔骨髓来源内皮祖细胞分离培养及鉴定

摘要

Objective To investigate how to isolate,culture and identify the endothelial progenitor cells from rabbit bone marrow in order to get ready for subsequent experiments.Methods EPCs were obtained from in vitro culture of MNCs which were isolated with density gradient centrifugation from rabbit bone marrow.Process of cell growth was observed.Cultivated cells were identified by immunofluorescence and cell chemical fluorescence detection.Results After 10 days cultured, EPCs were double positive for DilacLDL uptaking and FITC-UEA-1 binding and expressing CD34 and CD31 related antigen.Conclusion The mononuclear cells isoiated from rabbit bone marrow by gradient centrifugation can be induced into endothelial progenitor cells in vitro.The svstem of cell culture can be applied to subsequent experiments in cell transplantation.%目的 研究新西兰大白兔骨髓来源内皮祖细胞(EPC)的分离培养和鉴定方法,证实骨髓是获得内皮祖细胞的理想来源之一,为后续研究提供材料准备.方法 取兔骨髓,密度梯度离心法分离单个核细胞(MNC),置于DMEM-L培养基中培养,动态观察细胞生长过程,采用免疫荧光染色和细胞荧光化学法鉴定培养的细胞.结果 培养10 d的EPC能吞噬ac-LDL并与凝集素UEA-1相结合,同时表达CD34和CD31相关抗原.结论 密度梯度离心法从兔骨髓中分离的MNC在一定条件下,能够分化为具有内皮祖细胞特征的细胞,为后续的实验研究提供了细胞来源.