【摘要】 目的:分离培养兔脂肪间充质干细胞,探讨兔脂肪间充质干细胞在体外向神经细胞分化的能力。方法:实验于2002-09/2005-09在陕西省干细胞工程技术研究中心完成。选取清洁级新西兰大白兔5只,采用Ⅰ型胶原酶消化兔腹股沟皮下脂肪组织,分离培养脂肪间充质干细胞。取生长状况良好的第2,5,10代细胞,绘制生长曲线,计算细胞群体倍增时间。采用免疫组化方法对脂肪间充质干细胞表面分子标志[CD14,CD29,CD34,CD44,CD45,CD105及人白细胞抗原DR亚类(群)]进行鉴定,并采用1,2.5,5,10mmol/L不同浓度的β-巯基乙醇检测脂肪间充质干细胞向神经细胞分化的能力。结果:①脂肪间充质干细胞生长曲线分析:第2,5,10代细胞体外传代培养的潜伏期约为24～48h,传代培养细胞的对数增殖期为6d,接种后第9天进入平台期。在细胞生长的对数期,脂肪间充质干细胞的倍增时间为64.12h。②脂肪间充质干细胞分子特征的鉴定结果:第2～5代细胞90%以上CD29,CD44,CD105呈阳性表达,而不表达CD14,CD34,CD45及人白细胞抗原DR亚类(群)等表面分子标记。③脂肪间充质干细胞向神经细胞分化情况:体外能够分化为神经细胞。结论:兔脂肪组织可分离培养出脂肪间充质干细胞,并能够向神经细胞分化,有望成为组织工程的种子细胞来源。更多还原

【Abstract】 AIM: To isolate and culture mesenchymal stem cells from rabbit inguinal fat pads and identify the neurogenic differentiation of the adipose tissue-derived mesenchymal stem cells in vitro. METHODS: The experiment was conducted in Shaanxi Center of Stem Cell Engineering and Technology from September 2002 to September 2005. Inguinal fat pads of five health New Zealand White rabbits were digested with collagenase type I, and mesenchymal stem cells were derived by culture. Growth kinetics was determined and the population doubling time of these cells was calculated at passage 2, 5, 10. Molecular characteristics of these cells were identified by immunohistochemistry staining in CD14,CD29,CD34,CD44,CD45,CD105 and HLA-DR, and also its neurogenic potential under 1,2.5,5,10 mmol/L β-Me in vitro was identified. RESULTS: ①Analysis of growth curve of mesenchymal stem cells: The latency of cells at passage 2, 5, 10 was 24-48 hours, and the proliferative phase of cultured cells was 6 days, and then the cells entered the platform phase at the 9th day. The population doubling time of adipose tissue-derived mesenchymal stem cells was about 64.12 hours. ②Identification of characteristics of these cells: Over 90% rabbit adipose tissue-derived mesenchymal stem cells of passage 2-5 could express CD29, CD44, CD105 with positive expression, while did not express CD14, CD34, CD45 or HLA-DR. ③Differentiation of adipose tissue-derived mesenchymal stem cells into neurocytes: They had the ability to differentiate into neurocytes. CONCLUSION: Adipose tissue-derived mesenchymal stem cells can be isolated from rabbit adipose tissues, and have the potential to differentiate into neurocytes. It may be a source for tissue engineering.更多还原