摘要
背景:旨在从大鼠外周血及骨髓中提取内皮祖细胞,培养晚期内皮祖细胞,为干细胞移植治疗或通过内皮祖细胞联合基因治疗使内皮祖细胞高表达血管新生诱导因子,达到促进缺血性脑血管病血管新生的目的。目的:从大鼠骨髓及外周血中分离出内皮祖细胞并对其进行鉴定。方法:使用密度梯度离心及贴壁筛选法从大鼠骨髓和外周血中分离获得单个核细胞,进行诱导培养,观察并记录贴壁细胞的生物学特征;选取内皮祖细胞特异性表面标志CD133、CD34、KDR对原代细胞进行免疫荧光检测,利用流式细胞学技术检测KDR、CD34表达,并通过吞噬功能实验进一步鉴定培养细胞。结果与结论:大鼠骨髓和外周血能够分离获得早晚期内皮祖细胞;贴壁细胞免疫荧光检测CD34、CD133、KDR表达阳性;流式细胞学检测CD34、KDR表达阳性;贴壁细胞能够吞噬ac-LDL,结合UEA-1。实验成功从大鼠骨髓及外周血中分离出内皮祖细胞;并获得了增殖活性强的晚期内皮祖细胞,找到了更好的成血管干细胞的种子来源。
BACKGROUND： Our experiments intended to obtain bone marrow endothelial progenitor cells from rat peripheral blood and bone marrow, and culture the late endothelial progenitor cells to perform the stem cells transplantation or endothelial progenitor cells combined with gene therapy in order to highly express the angiogenic factors, thus to promote the angiogenesis after ischemic oerebrovascular disease. OBJECTIVE： To isolate and identify the endothelial progenitor cells from rat bone marrow and peripheral blood. METHODS： Mononuclear cells were obtained by using density gradient centrifugation and adherence screening method from bone marrow and peripheral blood of rats. Then the cells were induced, and the biological characteristics of the adherent cells were observed and recorded. Endothelial progenitor cells specific surface markers CD133, CD34 and KDR were selected and detected by immunofluorescence. FI0w cytometry was used to detect the expression of KDR and CD34. Phagocytosis experiment was carried out to further identify the cells. RESULTS AND CONCLUSION： Endothelial progenitor cells can be isolated from bone marrow and peripheral blood of rats; attached cells were positive for CD34, KDR and CD133 through immunofluorescence; cells were positive for CD34 and KDR by flow cytometry; cells could swallow acetylated-low-density lipoprotein and bind with Ulex europaeus agglutinin 1. The endothelial progenitor cells were isolated from bone marrow and peripheral blood of rats successfully and the late endothelial progenitor cells that have strong proliferative activity were obtained. A better source of stem cells was found to form blood vessels.
出处
《中国组织工程研究》
CAS
CSCD
2013年第6期1056-1063,共8页
Chinese Journal of Tissue Engineering Research
关键词
干细胞
干细胞培养与分化
内皮祖细胞
骨髓
外周血
血管新生
缺血性脑卒中
干细胞图片文章
stem cell
stem cell culture and differentiation
endothelial progenitor cells
bone marrow
peripheral blood
angiogenesis
cerebral ischemia stroke
stem cell photographs-containing paper
