海南医科大学学报2025，Vol.31Issue(14)：1050-1056,7.DOI:10.13210/j.cnki.jhmu.20250425.003
半悬浮组织器官培养法提取原代大鼠脂肪间充质干细胞及其鉴定
Semi-suspension organ culture method for the separation and culture of primary rat fatty mesenchymal stem cells and their identification
摘要
Abstract
Objective:To extract primary rat adipose-derived mesenchymal stem cells(AD-MSCs)using the semi-suspension organ culture method,providing sufficient and practical seed cells for regenerative medicine therapies and tissue engineering re-search.Methods:Inguinal and epididymal fat pads were aseptically isolated from 3-week-old SD rats.After disinfection,rinsing,and mincing,the tissues were digested with 0.1%collagenase Ⅱ for 30 minutes.The volume of low-glucose DMEM complete cul-ture medium was precisely controlled for semi-suspension primary culture.When confluence reached 90%,cells were passaged.The cells were further characterized by morphological observation,flow cytometry for surface markers(CD molecules),and adip-ogenic/osteogenic differentiation assays.Results:After the original inoculation and culture for 6-8 days,elongated spindle-shaped adherent cells grew to confluence,forming a dense"fibroblast-like"vortex pattern.Cells morphology was maintained through pas-sage 8 without significant"thickening"or differentiation.Flow cytometry of passage 4 cells revealed high CD90(97.3%),CD73(96.7%),and CD44(84.1%)expression,and low CD34(0.4%),CD45(8.1%),and CD11b/c(3.6%)expression.Adipogen-ic and osteogenic differentiation assays were positive.Conclusion:The semi-suspension organ culture method successfully isolates primary rat AD-MSCs.关键词
脂肪间充质干细胞/半悬浮组织器官培养法/原代培养/干细胞表面标志物/三系分化潜能Key words
Adipose-derived mesenchymal stem cells/Semi-suspension organ culture method/Primary culture/Surface markers of stem cells/Trilinear differentiation potential分类
医药卫生引用本文复制引用
唐元瑜,池茗,李柏霖,林志美,赖素玉,马华根..半悬浮组织器官培养法提取原代大鼠脂肪间充质干细胞及其鉴定[J].海南医科大学学报,2025,31(14):1050-1056,7.基金项目
This study was supported by the National Natural Science Foundation of China(81072714)and Natural Science Foundation of Fujian Province(2017J01545) 国家自然科学基金项目(81072714) （81072714）
福建省自然科学基金项目(2017J01545) （2017J01545）
