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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。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-tubulin蛋白表达阳性。脂肪诱导后油红O染色阳性。结论脐血中可分离出CD44+细胞,体外诱导可向神经、骨和脂肪方向分化。
Objective To isolate and culture human umbilical cord blood CD44 + cells and induce the differentiation into bone, nerve and adipose tissue. Methods CD44 + cells were isolated from cord blood by immunomagnetic beads after screening. Cells were osteogenic, respectively, into neural and adipogenic differentiation. Morphological characteristics were observed, flow cytometry was used to detect cell cycle and immunogenicity. Immunohistochemistry and Western blotting were used to detect the expression of cell differentiation protein. Results Human umbilical cord blood CD44 + cells adherent growth, long fusiform, cell proliferation rapidly after passage. Cell phenotypes were CD44 +, CD29 +, CD166 +, CD34-. Alkaline phosphatase staining was positive after osteogenic induction, and Von-Kossa staining suggested calcified nodule formation. Nestin, NSE and GFAP were positive after induction of neurotransmitter; Tau and β-tubulin protein were positive. Oil red O staining after fat induction. Conclusion CD44 + cells can be isolated from umbilical cord blood and induced in vitro to differentiate into nerve, bone and adipose tissue.