兔骨髓间充质干细胞的分离培养及肝内示踪  

Isolation,Culture and Hepatic Tracking of Rabbit Marrow Mesenchymal Stem Cells

作　　者：张芙蓉[1] 屠慧渭[1] 钱波[1] 孙何[1] 金立方[1] 

机构地区：[1]绍兴文理学院生命科学学院,浙江绍兴312000

出　　处：《动物医学进展》2011年第11期65-69,共5页Progress In Veterinary Medicine

基　　金：浙江省自然科学基金(Y2110911);浙江省大学生科技创新活动基金(2011R426010)

摘　　要：通过分离培养兔骨髓来源的间充质干细胞(MSCs),示踪其肝内移植命运,为MSCs的细胞治疗肝脏疾病提供理论依据和试验支持。采取全骨髓贴壁培养法分离培养兔骨髓MSCs,利用携带增强型绿色荧光蛋白(EGFP)的慢病毒载体感染MSCs,选择最优转染效率,并移植入经D﹣氨基半乳糖诱导的急性/亚急性肝衰竭受体兔体内,荧光显微镜下直接观察EGFP阳性细胞在肝内分布。分离出兔骨髓来源的MSCs,在体外可稳定传代,并具有成骨和成脂的分化潜能;慢病毒可高效感染MSCs;移植后第3、7、15天在肝功能衰竭的受体兔肝组织内可见GFP阳性细胞。携带EGFP的慢病毒载体具有高效转染兔MSCs的能力,并可作为报告基因简单有效示踪MSCs的肝内分布。To evaluate the feasibility for cell therapy effect for liver diseases,EGFP expressing rabbit bone marrow mesenchymal stem cells（MSCs） were isolated,cultured and transplanted into liver for tracking EGFP positive MSCs.The MSCs were isolated from rabbit bone marrow by adherent culture.The MSCs were infected by lentivirus carrying enhanced green fluorescent protein（EGFP）,and then transplanted into D-galactosamine-induced acute/subacute hepatic failure rabbits.The liver samples were collected and observed for EGFP positive cells under fluorescence microscope.The MSCs were isolated from adult rabbit bone marrow with potential to differentiation into bone and fat cells.They had robust proliferation capacity and were efficiently infected by lentivirus carrying EGFP.On day 3,7 and 15 after transplantation,GFP-positive cells were found in the livers of recipient.Thus,EGFP can be used as a simple and effective method to trace MSCs distribution in vivo.

关 键 词：骨髓间充质干细胞 兔 分离 示踪 增强型绿色荧光蛋白 

分 类 号：Q813.11[生物学—生物工程]