中国组织工程研究与临床康复2011，Vol.15Issue(1)：29-32,4.DOI:10.3969/j.issn.1673-8225.2011.01.006
SD大鼠脂肪源间充质干细胞分离培养特性及影响因素
Isolation and culture characteristics and influential factors of Sprague Dawley rat adipose tissue-derived mesenchymal stem cells
摘要
Abstract
BACKGROUND: Adipose tissue-derived mesenchymal stem cells (AD-MSCs) could be obtained easily with abundant quantities;the fat cells left in the donation area have the capacity of proliferation and the injury is pretty mild. Therefore, AD-MSCs have been an ideal source of seed cells for tissue engineering. However, the isolation and culture of AD-MSCs still remain difficulties.OBJECTIVE: To explore the method for isolation and culture of AD-MSCs and observe their growing properties.METHODS: The adipose tissues were obtained from the inguinal groove, shoulder and abdominal cavity of Sprague Dawley rats aged 4 weeks and 10 months. Following washes and centrifugation, specimens were incubated in cell culture flask, and placed in 5％CO2 incubator. The liquid was replaced at 24, 48 and 72 hours. At a fusion of 80％ , specimens were digested with 0.25％ trypsogen-EDTA and passaged. Effects of age, gender, and the region of collecting adipose tissue, collagenase Ⅰ concentration and digestion time, and first time of liquid change on the culture of AD-MSCs were analyzed.RESULTS AND CONCLUSION: Primary AD-MSCs presented spindle or multangular shape. While AD-MSCs passaged and cultu red longer, the newborn AD-MSCs, with polygon shape in clusters, joined to pieces and stretched into spindle or multangular shape gradually. A few fat cells appeared after 48 hours in culture and the cell number increased when the culture time was prolonged. However, about 90％ of the isolated cells still kept the shape of AD-MSCs. The AD-MSCs could be isolated successfully from the fat harvested from the 4-week female Sprague Dawley rat abdominal cavity, and be passaged stably in vitro.The Sprague Dawley rats' age, sex, collection site of adipose tissue, the concentration of collagenase Ⅰ and its digestion time,as well as the time of the first change of medium have an impact on the isolation and cultu re of AD-MSCs.关键词
脂肪源间充质干细胞/细胞来源/分离培养/组织修复/生物工程分类
医药卫生引用本文复制引用
侯凯,李梅,杨逸昆,李纯,李金茹,黄德清..SD大鼠脂肪源间充质干细胞分离培养特性及影响因素[J].中国组织工程研究与临床康复,2011,15(1):29-32,4.基金项目
2008年天津市应用基础及前沿技术研究计划项目(08JCYBJC27100),课题名称:脂肪源干细胞植入纳米纤维支架构建组织工程化肌腱.2007年天津市卫生局科技基金面上项目(07KR01):细胞因子诱导脂肪源干细胞强化组织工程化肌腱. （08JCYBJC27100）
