氯甲基苯甲酰胺标记大鼠肾脂肪囊来源脂肪间充质干细胞的可行性  

Feasibility of chlormethyl-benzamidodialkylcarbocyanine labeling rat kidney fat capsule adipose-derived mesenchymal stem cells

作　　者：訾利影[1] 郑红光[2] 张德伟[2] 梅煜明[2] 

机构地区：[1]辽宁医学院沈阳军区总医院研究生培养基地,辽宁省锦州市121011 [2]解放军沈阳军区总医院,辽宁省沈阳市110016

出　　处：《中国组织工程研究》2012年第19期3520-3524,共5页Chinese Journal of Tissue Engineering Research

基　　金：2011年辽宁省攻关课题~~

摘　　要：背景:氯甲基苯甲酰胺被认为是目前较为稳定的细胞荧光标记物质。目的:观察荧光染料氯甲基苯甲酰胺标记肾脂肪囊来源脂肪间充质干细胞的标记效果及其对细胞形态、生长及增殖能力等生物学特性的影响。方法:采用胶原酶消化法分离、培养SD大鼠肾脂肪囊来源脂肪间充质干细胞,以氯甲基苯甲酰胺细胞标记液标记第3代肾脂肪囊来源脂肪间充质干细胞。结果与结论:荧光显微镜下观察氯甲基苯甲酰胺标记的肾脂肪囊来源脂肪间充质干细胞呈红色荧光,细胞标记率达95%以上,标记前后细胞形态、生长及增殖等方面无明显变化。经多次传代培养后,红色荧光细胞数量及强度有所下降,但细胞传代至第8代,氯甲基苯甲酰胺标记后的第15天细胞标记率仍可达50%以上。表明氯甲基苯甲酰胺可有效标记肾脂肪囊来源脂肪间充质干细胞,且方法简单,标记率高,对细胞无毒性,不影响细胞生物学特性。BACKGROUND： Chlormethyl-benzamidodialkylcarbocyanine （CM-DiI） has been considered a relatively stable lipophilic fluorescent dye. OBJECTIVE： To investigate the labeling effects of CM-Dil for kidney fat capsule adipose-derived mesenchymal stem cells and its effects on cell morphology, growth and proliferation. METHODS： Sprague-Dawley rat kidney fat capsule adipose-derived mesenchymal stem cells were isolated by collagenase digestion. Passage 3 kidney fat capsule adipose-derived mesenchymal stem cells were labeled by CM-Dil. RESULTS AND RESULTS： Under fluorescent microscope, CM-Dil-labeled kidney fat capsule adipose-derived mesenchymal stem cells exhibited red fluorescence, with a labeling rate of over 95%. There were no obvious changes in cell morphology, growth and proliferation before and after labeling. After several passages, the number and staining intensity of cells with red fluorescence were decreased. At 15 days after CM-Dil labeling, over 50% of passaged cells were labeled. These suggest that CM-Dil can effectively label fat capsule adipose-derived mesenchymal stem cells and this method is easy-to-operate, with a high labeling rate, produces no cytotoxicitry and does not influence cell biological characteristics.

关 键 词：氯甲基苯甲酰胺(CM-DiI) 肾脂肪囊 脂肪间充质干细胞 标记 大鼠 

分 类 号：R394.2[医药卫生—医学遗传学]