人脐血CD44细胞分离培养与分化潜能初探.docVIP

PAGE 人脐血CD44+细胞分离培养与分化潜能初探 马云胜1 秦书俭2 穆长征1（[基金项目]：辽宁省教育厅基金（2008403）；辽宁省自然科学基金；辽宁省教育厅创新团队项目（2006T062）。[作者简介]： [基金项目]：辽宁省教育厅基金（2008403）；辽宁省自然科学基金；辽宁省教育厅创新团队项目（2006T062）。 [作者简介]：马云胜（1976-），男，辽宁省锦州市人，讲师，博士研究生；主要研究方向：干细胞多向分化潜能与组织修复。 通讯作者：穆长征（1962-），男，教授，硕士生导师，方向：干细胞治疗与组织修复，E-mail: HYPERLINK mailto:yunshengma@126.com yunshengma@126.com。 Isolation, culturing and differentiation CD44+ cells from human cord blood MA Yun-sheng1, QIN Shu-jian2, MU Chang-zheng1 1 Department of Histology and Embryology, Liaoning Medical College. 2 Department of Anatomy, Liaoning Medical College [Abstract] Objective Isolate and culture CD44+ cells from human cord blood, study the charicristics of osteogenesis, neurogenesis and adipogenesis, and select seed cells for tissue engineering. Methods The CD44+ cells were isolated by immunomagnetic beads from cord blood. P2 cells were to study the ability of osteogenesis, neurogenesis, adipogenesis. The cells morphology were observed. Then the cell cycles, immunogenicity and phonotype were tested by flowing cytometry. The ability of differentiation was tested by immunohistochemistry and western blot. Results The cord blood CD44+ cells presented spindle shape. The phenotype was CD44+, CD29+, CD166+, CD34-. In osteogenic experiment the cells’ alkaline phosphatase(ALP) was positive, and hydroxyapatite nodules were formed by Von-Kossa staining. Nestin, NSE and GFAP were positive in neural inducement experiment. red O staining was positive in adipose differentiation experiment. Conclusion The cord blood CD44+ cells could differentiate into osteoid , liparoid and neural-like cells in vitro. Key word: cord blood; CD44+; differentiation; seed cells [摘要] 目的 分离培养人脐静脉血CD44+细胞，并且诱导其向骨、神经和脂肪组织分化，为组织工程提供种子细胞来源。 方法 采集的脐血用免疫磁珠阳性筛选法分离CD44+细胞后进行培养。细胞分别行成骨、成神经和成脂诱导分化。观察细胞形态学特征，流式细胞仪检测细胞周期、免疫原性。免疫组化及蛋白印记法检测细胞分化蛋白表达。结果 人脐血CD44+细胞贴壁生长，长梭形，传代后细胞增殖迅速。细胞表型为CD44+，CD29+,CD166+，CD34-。在成骨诱导后碱性磷酸酶染色阳性，Von-Kossa染色提示钙化结节形成。成神经诱导后Nestin, NSE, GFAP阳性；Tau和β-tubu