人脐血CD44细胞多向分化潜能研究.doc

人脐血CD44+细胞多向分化潜能研究 马云胜1 秦书俭2 穆长征1（辽宁医学院组织胚胎学教研室,2辽宁医学院解剖学教研室, 辽宁 锦州121001） Multipotential differentiation characteristics of 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, Jinzhou China 121001. [Abstract] Objective To study the charicristics of osteogenesis, neurogenesis, and adipogenesis of CD44+ cells from human cord blood, 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 探讨来源于人脐静脉血CD44+细胞向骨、神经和脂肪组织分化的能力，为异体细胞移植治疗提供种子细胞来源。 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。P2代细胞分别行成骨、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-tubulinШ蛋白表达阳性。脂肪诱导后油红O染色阳性。结论 脐血CD44+细胞能够在体外诱导分化为成骨细胞、成脂细胞。 ［关键词］ 脐血；CD44+；诱导分化；种子细胞 [中图分类号] R329.21 [文献标识码] A [文章编号] 0 引言 （mesenchymal stem cell，MSC）脐带血中富含多种亚群，。造血干细胞CD34+细胞，有研究表明CD34+细胞中有亚群，细胞呈贴壁生长且在一定条件下可向骨，神经和脂肪方向分化[]。我们在研究中应用免疫学原理从脐血中提纯CD44+细胞发现脐