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

摘要

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. n 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. n 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) × 106 cells. There were (1.60 ± 0.59) × 106 of PSCs, (0.18 ± 0.04) × 106 of pericytes and (1.42 ± 0.57) × 106 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). n 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.%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。n　　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。n　　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）。n　　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。