2. 牛脂肪间充质干细胞的分离培养与鉴定.任宇 .pdfVIP

生 物 工 程 学 报 Chin J Biotech 2010, December 25; 26(12): 1645−1651 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn ©2010 CJB, All rights reserved. 干细胞专栏 牛脂肪间充质干细胞的分离、培养与鉴定 1 1 1,2 1 1 1 任宇 ，吴海青 ，马玉珍 ，仓明 ，王瑞 ，刘东军 1 010021 2 010017 摘 要: 为了给组织工程提供种子细胞，对牛间充质干细胞 (Adipose-derived stem cells ，ADSCs) 进行体外分离培养。 首先应用胶原酶消化法分离牛ADSCs ，进行体外培养、连续传代，并观察细胞的形态变化，通过细胞计数绘制生长曲 线，细胞压片进行染色体分析，采用细胞免疫荧光化学方法检测细胞表面标记，利用成骨分化和成脂分化检测其分化 能力。结果显示牛ADSCs 体外培养时细胞形态呈成纤维细胞样，增殖稳定；Vimentin 、CD49d 、CD13 表达呈阳性， CD34 表达呈阴性；成骨诱导条件下的细胞碱性磷酸酶活性高，茜素红染色呈阳性；成脂诱导条件下细胞周围脂滴明显， 油红-O 染色呈阳性。结果证明牛 ADSCs 体外生长稳定、增殖速度快、定向分化能力强，简易的体外分离培养及诱导 方法为其在组织工程中的应用奠定了基础。 关键词: 脂肪间充质干细胞，体外分离培养，细胞鉴定，细胞分化 Isolation, cultivation and identification of adipose-derived stem cell in bovines 1 1 1,2 1 1 1 Yu Ren , Haiqing Wu , Yuzhen Ma , Ming Cang , Rui Wang , and Dongjun Liu 1 Key Laboratory of Ministry of Education of China for Mammal Reproduction Biology and Biotechnology, Inner Mongolia University, Huhhot 010021, China 2 Inner Mongolia Hospital , Hohhot 010017, China Abstract: To obtain bovine adipose-derived stem cells (ADSCs), bovine ADSCs were digested in collagenase type I solution. The growth curve of ADSCs was checked by cell counting. Chromosome analysis was checked. The molecular markers of ADSCs were detected with immunofluoresence staining. The morphology of ADSCs was identical to fibroblast 1ike and the cells showed active proliferative ability. Vimentin, CD49d and CD13 antigens were detected, but CD34 antigen was negative. Alka