CM-Dil体外标记脂肪基质干细胞及其在脂肪移植裸鼠模型体内示踪研究  

CM-Dil labeling on adipose-derived mesenchymal stem cells in vitro and its tracking in fat graft tissues in nude mouse model

作　　者：吕成鹏 倪华 马锐 李龙 刘斌 刘彦普 L Chengpeng;NI Hua;MA Rui;LI Long;LIU Bin;LIU Yanpu(Department of Stomatology,Shaanxi Provincial People’s Hospital,Xi’an 710068,China;Department of Oral and Maxillofacial Surgery,School of Stomatology,Airforce Military Medical University;Tissue Engineering Center,School of Stomatology,Airforce Military Medical University;Department of Anesthesiology,Xi’an Children’s Hospital;Department of Stomatology,Shenzhen Nanshan District Shekou People’s Hospital;Laboatory animal Center,School of Stomatology,Airforce Military Medical University)

机构地区：[1]陕西省人民医院口腔科,西安710068 [2]空军军医大学口腔医院口腔颌面外科 [3]空军军医大学口腔医院组织工程研发中心 [4]陕西省西安市儿童医院麻醉科 [5]广东省深圳市南山区蛇口人民医院口腔科 [6]空军军医大学口腔医院动物实验中心

出　　处：《山西医科大学学报》2020年第3期237-241,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81070820);陕西省科学技术厅社会发展领域项目资助课题(2019SF-161)。

摘　　要：目的探讨CM-Dil对脂肪基质干细胞(ADSCs)的体外标记及其在脂肪移植裸鼠模型的体内示踪。方法采用贴壁培养法分离、纯化、扩增ADSCs。取第3代细胞进行CD29、CD31免疫组化染色,用流式细胞仪检测细胞阳性率;对第3代细胞进行成骨、成脂诱导,鉴定其成骨成脂多向分化潜能。CM-Dil体外标记ADSCs,并用荧光显微镜检测。6周龄雌性BALB/c裸鼠制备脂肪移植裸鼠模型,将标记CM-Dil的ADSCs脂肪移植复合体移植入裸鼠背部皮下,移植后4周取脂肪移植物制备冰冻切片,荧光显微镜观察ADSCs的分布情况。结果脂肪组织中分离培养的ADSCs呈梭形生长,CD29表达阳性细胞比例为84.52%±4.91%,CD31表达阳性细胞比例为2.13%±0.95%,ADSCs能够分化为成脂细胞和成骨细胞。荧光显微镜显示,CM-Dil标记的ADSCs在裸鼠脂肪移植中表达阳性。结论CM-Dil能够体外标记ADSCs,可用于ADSCs在脂肪移植裸鼠模型的体内示踪,ADSCs可能参与了脂肪移植后的组织重建。Objective To investigate the cell labeling of CM-DiI on the adipose-derived mesenchymal stem cells(ADSCs)in vitro and its tracking in fat graft in nude mouse.Methods ADSCs were isolated,purified and amplified by adherent culture.The third-generation cells were taken for CD29 and CD31 immunohistochemical staining,and the positive rate was detected by flow cytometry.The third-generation cells were subjected to osteogenesis and adipogenesis induction to identify their osteogenic and adipogenic multipotential differentiation potential.Then ADSCs were labeled in vitro by CM-DiI,and were observed under the fluorescence microscope.The 6-week-old female BALB/c nude mouse was used to prepare a fat transplantation model.ADSCs labeled with CM-DiI were transplanted subcutaneously into the back of nude mice.The fat grafts were taken 4 weeks after transplantation to prepare frozen sections.The distribution of ADSCs was observed with a fluorescence microscope.Results ADSCs isolated from adipose tissue grew spindle-shaped.The percentages of CD29 positive cells and CD31 positive cells were 84.52%±4.91%and 2.13%±0.95%,respectively.ADSCs were able to differentiate into adipocytes and osteoblasts.Fluorescence microscopy showed that CM-Dil labeled ADSCs were positive in fat transplantation in nude mice.Conclusion CM-Dil is capable of labeling ADSCs in vitro,and can be used to track ADSCs for fat transplantation experiment in vivo.ADSCs may be involved in tissue reconstruction after fat transplantation.

关 键 词：CM-Dil 细胞示踪 脂肪基质干细胞 脂肪移植 裸鼠 

分 类 号：R318[医药卫生—生物医学工程]