血管周围干细胞在大鼠脂肪组织中的含量及其体外扩增后比例的变化  

Proportion of perivascular stem cells(PSCs) in adipose tissue of rat and the content changes of PSCs in ADSCs

作　　者：徐峰[1] 刘舒云[1] 彭江[1] 卢世璧[1] 孙逊[1,2] 尹合勇 孙振[1] 余晓明[1] 肖波[1] 张金鑫[1,3] 袁玫[1] 许文静[1] 郭全义[1] 

机构地区：[1]解放军总医院骨科研究所,北京100853 [2]南开大学医学院,天津300071 [3]佳木斯大学研究生院,黑龙江154007

出　　处：《中国医药生物技术》2015年第2期97-101,共5页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(81472092);国家高技术发展研究计划(863计划)(2012AA020502)

摘　　要：目的探索大鼠脂肪组织中血管周围干细胞(PSCs)在脂肪组织细胞中所占的比例,以及体外培养扩增后的大鼠脂肪间充质干细胞(ADSCs)中PSCs比例的变化,为组织工程自体细胞移植修复骨和软骨损伤的实验研究提供可能的新型种子细胞。方法取大鼠的脂肪组织用II型胶原酶消化得到血管基质成分(SVF)并且体外培养扩增大鼠ADSCs至P3代,用细胞计数仪及流式细胞仪检测SVF细胞密度、活细胞比例、PSCs含量,检测ADSCs中PSCs的含量并与CD45–SVF细胞中PSCs含量进行比较。结果经细胞计数仪分析,每只大鼠约3 ml脂肪组织消化所得SVF中活细胞比例为(69.31±6.57)%,可收获活细胞(40.69±6.81)×106个。经流式细胞仪检测,每只SD大鼠3 ml脂肪组织中平均可以得到PSCs活细胞为(1.60±0.59)×106个,血管外周细胞为(0.18±0.04)×106个,血管外膜细胞为(1.42±0.57)×106个。而体外平面培养扩增的P3代ADSCs中,PSCs及各细胞亚群的比例并没有随着细胞的扩增而改变(P>0.05)。结论大鼠脂肪组织中含有PSCs,可以直接通过流式分选获取大量PSCs,不进行体外扩增即可望满足骨和软骨组织工程种子细胞的需要。Objective To explore the proportion of PSCs in adipose tissue cells of rat and the proportion changes of PSCs in ADSCs which were cultured in vitro. This research aims to provide potential new seed cells for autologous cell transplantation to repair the injuries of bone and cartilage in tissue-engineering. Methods Stromal vascular fraction（SVF） was isolated by collagenase type II digestion from the SD rat adipose tissue and ADSCs was cultured in vitro until P3. The cell density of SVF cells, the proportion of living cells and the proportion of PSCs in SVF were measured by the cell counter and flow cytometry（FCM）. The proportion of PSCs in ADSCs was compared with that in CD45–SVF cells. Results The proportion of live cells in SVF isolated from each 3 ml of rat adipose tissue was（69.31 ± 6.57）%, reaching（40.69 ± 6.81） × 10^6 cells. There were（1.60 ± 0.59） × 10^6 of PSCs,（0.18 ± 0.04） × 10^6 of pericytes and（1.42 ± 0.57） × 10^6 of adventitial cells obtained from each 3 ml of rat adipose tissue by the flow cytometry sorting. In the in vitro cultured P3 of ADSCs, the proportions of PSCs and cell subsets were not significantly changed（P〉0.05）. Conclusion This research suggests that there are enough PSCs in the adipose tissue of rat and the content of PSCs from flow cytometric sorting in the tissue may meet the needs of seed cells for bone and cartilage tissue-engineering without in vitro culture.

关 键 词：血管周围干细胞 脂肪间充质干细胞 流式细胞术 

分 类 号：R687[医药卫生—骨科学] R318.08[医药卫生—外科学]