| 摘要: |
| 【目的】分离、培养奶山羊骨髓间质干细胞（MSCs），探讨其生物学特性，为心血管、神经退化,尤其是不育症患者的细胞移植治疗及相关细胞发育分化研究奠定基础。【方法】 采用贴壁培养法进行MSCs的分离和培养，通过形态学观察、生长曲线测定、免疫细胞化学染色和RT-PCR技术等进行检测；并采用免疫细胞化学法对自发分化的细胞及定向诱导分化的神经样细胞、心肌样细胞、生殖样细胞进行检测，计算生殖样细胞的阳性率。【结果】分离得到的MSCs为成纤维样细胞，其增殖能力强，无致瘤性，已连续传代至22代；其表达胚胎干细胞（ESCs）的表面标记Oct4、Nanog、C-myc和TERT；MSCs形成类胚体自发分化的细胞表达3胚层特异标志基因AFP、β-Ⅲ Tubulin和心肌α-actin；MSCs经β-巯基乙醇诱导，定向分化为表达Nestin和β-ⅢTubulin的早期神经样细胞,经5-氮胞苷（5-Aza）诱导，可定向分化为表达CT3及心肌α-actin的心肌样细胞，并出现肌管样结构，经维甲酸（RA）诱导，分化的细胞表达生殖细胞减数分裂前期的蛋白Vasa、Scp3和CD49f(α6整合素)等生殖细胞标记，其阳性率分别达到35.7%，24.0%和14.0%。【结论】自奶山羊骨髓分离得到MSCs，其具备间质干细胞的基本特性，并具有向神经细胞、心肌细胞、生殖细胞分化的潜能。 |
| 关键词: 奶山羊 骨髓 间质干细胞 神经细胞 心肌细胞 生殖细胞 |
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| 基金项目:国家自然科学基金项目（30972097）；中国博士后基金项目（20080431253）；奶山羊良种繁育及产业化技术体系建设项目（nyhyzx07-037）；江苏科技基础设施建设计划“科技公共服务平台”项目（BM2008146） |
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| Isolation and characterization of diary goat bone marrow derived mesenchymal stem cells |
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| Abstract: |
| 【Objective】The diary goat bone marrow-derived mesenchymal stem cells (MSCs) were isolated to investigate the characteristics to establish a model for cell differentiation and to provide new resources of cell therapy for neural and myocardial degenerative diseases,infertility and sterility.【Method】The cells were isolated and derived from dairy goat bone marrow by adherent culture,identified by morphology,growth curves,immunocytochemistry and RT-PCR.The positive rates of spontaneous differentiation cells were evaluated by immunocytochemical method.【Result】The isolated MSCs were fibroblast-like phenotype without-tumorigenicity and continuously passaged to 22;The markers of embryonic stem cells (ESCs) such as Oct4,Nanog,C-myc and TERT were identified.The spontaneously differentiated cells derived from embryoid bodies expressed markers of three embryonic germ layers.MSCs were induced to differentiate into early neural-like cells by β-mercaptoethanol induction and these cells were positive for Nestin and β-ⅢTubulin.Myocardial cells were obtained from MSCs induction by 5 aza 2′-deoxycytidine (5 Aza) and appeared myotube like cells,which were positive for CT3 and cardiac α-actin.Through RA induction,the differentiated cells expressed the specific markers of pre meiotic germ cells including Vasa,Scp3,CD49f and the positive rates were 35.7%,24.0% and 14.0% respectively.【Conclusion】These results demonstrated that typical mesenchymal stem cells were isolated from dairy goat bone marrow,possessed the characteristics of pluripotent stem cells, and had the potential of differentiation into neural,myocardial cells and even germ cells. |
| Key words: goat bone marrow mesenchymal stem cells (MSCs) neural cells myocardial cells germ cells |
