圈养孟加拉虎脂肪间充质干细胞的分离培养和生物学特性  

Isolation,Culture and Biological Characteristics Exploration of Adipose Mesenchymal Stem Cells of Captive Bengal Tiger

作　　者：冼伟杭 张天佑 赖健仪 王丙云 董贵信 陈志胜 陈胜锋 白银山 刘璨颖 张晖 计慧琴 XIAN Weihang;ZHANG Tianyou;LAI Jianyi;WANG Bingyun;DONG Guixin;CHEN Zhisheng;CHEN Shengfeng;BAI Yinshan;LIU Canying;ZHANG Hui;JI Huiqin(Collage of Life Science and Engineering,Foshan University,Foshan,528200,China;Guangdong Chimelong Group,Co.,Ltd.,Guangzhou,511430,China;Southern China Rare Wildlife Species Conservation Center,Zhuhai,519031,China)

机构地区：[1]佛山科学技术学院生命科学与工程学院,佛山528200 [2]广东长隆集团有限公司,广州511430 [3]广东华南珍稀野生动物物种保护中心,珠海519031

出　　处：《野生动物学报》2023年第4期807-814,共8页CHINESE JOURNAL OF WILDLIFE

基　　金：广东省长隆基金会项目(ZLPF2021002Z);广东省普通高校动物干细胞工程技术研究中心项目(2021GCZX006)。

摘　　要：圈养孟加拉虎(Panthera tigris tigris)是保护该濒危物种的有效途径,但限制性的圈养环境导致圈养虎的健康问题日益凸显,尤其是肾炎,当前的治疗方法预后效果较差。为分离获取、鉴定并培养具有治疗孟加拉虎肾炎治疗潜力的孟加拉虎脂肪间充质干细胞,通过采集孟加拉虎的脂肪组织,利用Ⅰ型胶原酶消化法分离脂肪间充质干细胞并进行传代培养;采用CCK8法测定P3、P6和P9代细胞活性并绘制生长曲线;采用PCR法扩增间充质干细胞表面特殊标记物基因;通过成脂肪和成骨诱导分化试验分别测定脂肪间充质干细胞的成脂与成骨诱导分化能力。结果发现:孟加拉虎脂肪间充质干细胞为长梭形,折光性较好,生长情况良好;P3、P6和P9代细胞生长曲线呈典型的“S”型,符合间充质干细胞生长曲线规律;PCR扩增得到CD44、CD90、CD105和CD29特异性表面标记物基因;诱导分化试验表明,脂肪间充质干细胞具有成脂成骨诱导分化能力。综上,分离培养获得的细胞符合间充质干细胞特征,表明成功分离出孟加拉虎脂肪间充质干细胞。虎脂肪间充质干细胞的成功分离将作为一种潜在的细胞治疗手段,可能为虎肾炎的治疗提供新的选择,有望在维持虎的健康状态和延长寿命方面产生积极的临床效果。Captive breeding of the Bengal tiger(Panthera tigris tigris)represents an effective approach for preserving this endangered species.However,the confined conditions of captivity cause health issues in tiger,with nephritis being a particular concern.Current treatment modalities for nephritis in captive tigers are plagued by limited efficacy.In order to isolate,identify and culture adipose mesenchymal stem cells of Bengal tiger for nephritis treatment,adipose tissue was collected,and adipose mesenchymal stem cells were isolated using type Ⅰ collagenase digestion method then followed by subsequent culture.The cell proliferative activities of P3,P6 and P9 generations were measured using CCK8 method and the growth curves were plotted.Specific marker genes of the mesenchymal stem cells surface protein were verified using PCR and the adipogenic and osteogenic differentiation ability of adipogenic mesenchymal stem cells was evaluated by inducing using specific media.The results showed that the Bengal tiger adipose mesenchymal stem cells exhibited an elongated spindle-like morphology with commendable refractivity and proliferation.The growth curves of P3,P6 and P9 generation cells showed a typical “S” shape,which were consistent with the growth curve of mesenchymal stem cells reported.In addition,surface marker genes CD44,CD90,CD105 and CD29 were successfully amplified by PCR.The induced differentiation experiment results showed that adipose mesenchymal stem cells displayed the ability to differentiate into adipogenic and osteogenic lineages.Based on these results,the cells obtained in this study showed characteristics consistent with previously reported mesenchymal stem cells,indicating that Bengal tiger adipose mesenchymal stem cells were successfully isolated.The effective isolation of tiger adipose mesenchymal stem cells holds promise as a potential cell-based therapeutic intervention for tiger nephritis,which may prove to have significant clinical benefits in terms of preserving the overall health and extending

关 键 词：孟加拉虎 肾炎 脂肪 间充质干细胞 再生疗法 生物学特性 

分 类 号：Q954.6[生物学—动物学]