悬浮法及酶消化法分离培养脂肪干细胞  

Isolation and culture of adipose-derived stem cells by suspension culture and enzyme digestion

作　　者：赵雅丽 李瑞方 熊丽娇[2,3] 曾治平 ZHAO Ya-li;LI Rui-fang;XIONG Li-jiao;ZENG Zhi-ping(Gannan Medical University;Department of General Medicine,the First Affiliated Hospital of Gannan Medical University;National Center for Geriatrics Clinical Research Gannan sub-center,Ganzhou,Jiangxi 341000)

机构地区：[1]赣南医学院 [2]赣南医学院第一附属医院全科医学科 [3]国家老年疾病临床医学研究中心赣南分中心,江西赣州341000

出　　处：《赣南医学院学报》2021年第10期984-987,共4页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：2018年赣南医学院校级课题(ZD201818)。

摘　　要：目的:了解脂肪干细胞(Adipose-derived stem cells,ADSCs)的获取及原代培养方法,明确体外定向分化条件,为进一步ADSCs移植实验提供基础。方法:取6周龄雄性大鼠,脱颈处死,在无菌条件下取大鼠腹股沟处脂肪组织,放入装有PBS的培养皿中,剔除肉眼可见的血管及筋膜,分别用酶消化法及悬浮组织块法培养、传代及鉴定,并比对两种方案的优缺势。结果:悬浮法和酶消化法培养得到的大鼠ADSCs形态均为梭形纤维样,生长迅速,生长曲线为典型的“S”形,两种方法获得的ADSCs经成骨诱导培养3周后茜素红染色均为阳性,成脂诱导培养3周后,油红染色也均为阳性。结论:大鼠提取的脂肪组织经两种培养方法均可定向诱导分化为ADSCs,并可稳定增殖传代,但悬浮法较酶消化法更便利,价格更低廉。Objective:To understand the methods of adipose derived stem cells(ADSCs)acquisition and primary cul⁃ture,and to clarify the conditions of directional differentiation in vitro,so as to provide the basic conditions for further transplantation of ADSCs.Methods:Six-week-old male rats were killed by stripping off their necks.The rat groin adi⁃pose tissue was taken under aseptic condition and put into a PBS petri dish to remove the visible blood vessels and fascia,the method of enzyme digestion and suspension culture were used to culture,subculture and identification,and the ad⁃vantages and disadvantages of two schemes were compared.Results:The morphology of rat ADSCs cultured by suspen⁃sion method and enzyme digestion method was fusiform,which grew rapidly and the growth curve was typical"S"shape.After 3 weeks of osteogenic induction culture,alizarin red staining was positive for ADSCs obtained by both method.Con⁃clusion:The adipose tissue of rats could be induced to differentiate into ADSCs by two methods,and could proliferate stably,but the suspension method was more convenient and cheaper than the enzyme digestion method.It is worthy of broader application.

关 键 词：脂肪干细胞 悬浮组织块法 酶消化法 大鼠 

分 类 号：R394.2[医药卫生—医学遗传学]