皮肤切口对皮下脂肪中脂肪间充质干细胞的影响

Effects of skin wound on adipose-derived mesenchymal stem cells harvested from subcutaneous adipose tissue in rats

目的:研究皮肤切口对皮下组织脂肪间充质干细胞(ADMSCs)的收获率、细胞形态、分化能力、增殖特性和表面标志的影响。方法:6只SD大鼠在左侧腹股沟做皮肤切口后缝合,和对照组6只大鼠同取右侧腹股沟脂肪垫分离培养ADMSCs。计算间质血管碎片细胞和贴壁细胞收获率,用这两种第4代ADMSCs进行成骨成脂诱导分化,绘制生长曲线并测定倍增时间。流式细胞术检测表面标志CD11b、CD29、CD45、CD49d、CD90和CD106的表达率。结果:皮肤切口组大鼠的间质血管碎片细胞和贴壁细胞收获率较对照组大鼠明显降低。来自两组大鼠的ADMSCs细胞形态上无明显差异,都能进行成骨成脂分化,生长曲线和细胞倍增时间无明显差异,表面标志CD11b、CD29、CD45、CD49d、CD90和CD106的表达率无明显差异。结论:皮肤切口能引起皮下脂肪组织的ADMSCs收获率降低,对获得的ADMSCs在形态、多向分化性质、增殖能力和表面标志方面无明显影响。

Objective To investigate the effects of skin wound on the morphology, yield,multiple differentiations, growth characteristics and cell surface markers of adipose-derived mesenchymal stem cells （ADMSCs） in rats. Methods 6 Sprague-Dawley rats were suffered skin incision in left inguen and the wounds were sutured. After 1 week, ADMSCs of injured rats and 6 normal rats from right inguinal fat pads were isolated and cultured. The yield of stromal vascular fraction cells and adherent cells were investigated. Both kinds of ADMSCs at passage 4 were induced to adipogenic and osteogenic differentiation. Growth kinetics was investigated and doubling times were determined. The phenotypical marker profile including CDllb, CD29, CD45, CD49d, CD90 and CD106 of ADMSCs grown for 4 passages was determined by flow cytometry. Results The yield of stromal vascular fraction cells and adherent cells from the injured rats is fewer than it from the normal rats. The morphological characterization of these ADMSCs were almost the same. Both kinds of ADMSCs could differentiate into adipogenic and osteogenic lineages. The growth curves and the mean population doubling time of these two kinds of ADMSCs showed no obvious differences. The frequency ofCD11b^＋,CD29^＋,CD45^＋,CD49d^＋,CD90^＋, and CD106^＋ ADMSCs derived from injured rats and normal rats were similar. Conclusion The yield of ADMSCs in injured rats is decreased, which may be influenced by skin wound. ADMSCs cultured from injured and normal rats are similar in multiple differentiations, growth characteristics and cell surface markers.