活体小鼠脂肪来源干细胞的获取与鉴定

Separation,extraction,culture and identification of adipose-derived stem cells from living mice

目的探究一种从活体小鼠身上提取脂肪来源干细胞(adipose-derived stem cells,ADSCs)的实验方法。方法自2020年3月至2020年7月,中国医科大学附属第一医院整形外科麻醉C57小鼠后从其腹膜外及腹股沟处分离脂肪组织,采用胶原酶消化,获得ADSCs。传至3代后,使用流式细胞仪分别检测其表面标志物表达情况,并对其进行诱导分化,验证其成脂肪,成骨分化的能力。结果从小鼠腹膜外及腹股沟部位可获得大量脂肪组织,且小鼠麻醉苏醒后生命体征平稳。ADSCs提取成功,14 d后观察小鼠生命体征平稳。流式细胞仪检测提示其CD29、CD44阳性,CD34阴性,成功对细胞进行成脂及成骨诱导,表明提取的ADSCs具有多向分化能力。结论本研究证实,可以从活体小鼠获取纯度较高的ADSCs且具有成脂肪、成骨分化能力。

Objective To explore an experimental method for extracting adipose-derived stem cells(ADSCs)from living mice.Methods From March 2020 to July 2020,C57 mice were anesthetized by the Plastic Surgery Department of the First Affiliated Hospital of China Medical University and adipose tissue was separated from the extraperitoneum and groin,and digested with collagenase to obtain ADSCs.After passing to the third generation,the expression of the surface markers was detected by flow cytometry,and the differentiation was induced to verify the ability of fat and osteogenic differentiation.Results A large amount of adipose tissue can be obtained from the mouse extraperitoneum and groin,and the vital signs of the mouse are stable after anesthesia.The ADSCs were successfully extracted.After14 days,the vital signs of the mice were observed to be stable.Flow cytometry showed that it was positive for CD29 and CD44,and negative for CD34.It successfully induced adipogenesis and osteogenic cells,indicating that the extracted ADSCs have multidirectional differentiation ability.Conclusion This study confirmed that ADSCs with high purity can be obtained from living mice and have the ability to differentiate into fat and osteogenic.