摘要
目的 :验证脐带血中内皮细胞前体细胞的存在 ,并研究这一前体细胞的分化发育过程中的影响因素。方法 :用淋巴细胞分离液分离出脐带血中的单个核细胞 ,在胶原包被的培养瓶中 ,用含 2 0 %小牛血清的MCDB131培养液培养 ,获得具有粘附性的细胞 ,并观察不同培养条件对这一细胞分化发育的影响。倒置相差显微镜下每日两次观察、记录。流式细胞分析所获细胞的CD34、CD14以及因子免疫组化检测Ⅶ因子表达。结果与结论 :脐带血中存在可粘附CD34-内皮细胞的前体细胞 ,小牛血清、小牛下丘脑提取物、胰岛素对这一细胞的生成及分化有促进作用 ,而地塞米松、次黄嘌呤则对这一细胞的生成及分化有抑制作用。免疫组化证实这一细胞Ⅶ因子表达阳性 ,培养两周的粘附梭形细胞可形成管状排列及血岛样结构。
AIM: To confirm the existence of the endothelial progenitor cells in human cord blood and to study its differentiation and development process. METHODS: The mononuclear cells in human cord blood were isolated using lymphocyte separation solution. Then the mononuclear cells were cltured in MCDB131 containing 20% fetal bovine serum. The effects of 5 μmol/L dexamethasone,the extract from bovine brain,insulin and hypoxanine on the proliferation and differentiation of the adherent cells were observed. The morphology of the adherent cells were examined twice daily by inverted phase contrast microscope. CD34 and CD14 expression were determined by FACS. Immunohistochemistry was used to confirm the expression of factor Ⅷ. RESULTS: The proliferative endothelial progenitor cells existed within the CD34 - adherent mononuclear cells of human cord blood. Dexamethasone and hypoxanine decreased the number of spindle-shaped cells and caudated cells. Bovine brain extract,insulin and FCS enhanced the number of spindle-shaped cells and caudated cells. CONCLUSION: The existence of endothelial progenitor cells within the CD34 - adherent monouclear cells of the human cord blood was observed and these cells were able to differentiate into endothelial-like cells in vitro.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2003年第11期1449-1452,共4页
Chinese Journal of Pathophysiology
