诱导时间对脂肪干细胞来源的类许旺细胞增殖率的影响  

Investigation effect of induction time on proliferation rate of induced Schwann-like cells from adipose derived stem cells

作　　者：黄昭伟 朱昭炜[1] 许澍洽[1] 林珣珣[1] 韩兵 刘祥厦[1] 许扬滨[1] Huang Zhaowei;Zhu Zhaowei;Xu Shiiqia;Lin Xunxun;Han Bing;Liu Xiangxia;Xu Yangbin(Department of Plastic Surgery,The First Affiluiled Hospital of Sun Yal-sen University,Guangzhou 510080,China)

机构地区：[1]中山大学附属第一医院整形外科,广州510080

出　　处：《中华显微外科杂志》2019年第2期150-154,共5页Chinese Journal of Microsurgery

基　　金：国家自然科学基金(81201546);广东省自然科学基金(2018A0303130217,2017A030310302);广东省医学科学技术研究基金(A2016018);广东省科技计划项目(2016A010103012).

摘　　要：目的观察比较诱导时间对脂肪干细胞(ADSC)向类许旺细胞(iSC)诱导分化时的增殖率的影响。方法2017年3月至2018年10月,选取健康成年雌性SD大鼠腹股沟脂肪分离培养ADSC,流式细胞仪检测ADSC标志阳性蛋白CD29、CD90及阴性蛋白CD45。iSC诱导液培养ADSC。通过S100和GFAP免疫荧光染色确认ADSC分化为iSC。诱导后第1、4、7、10、13、16和19天用光学显微镜观察细胞的形态变化,MTS法检测细胞增殖情况,Tunel染色检测细胞凋亡情况。将数据进行统计学分析。结果iSC均有S100、GFAP蛋白的表达。诱导第7天时细胞增殖率较诱导前明显减慢(A值分别为0.330±0.020和0.400±0.004,P<0.05),并与诱导时间负相关;第19天时,iSC增殖率低于诱导前的50%(A值分别为0.016±0.003和0.400±0.004,P<0.05)。iSC细胞凋亡较同期ADSC明显。结论ADSC诱导的iSC增殖率可能在诱导7d时活力最佳。Objective Comparison of induction time on the proliferation of induced adipose-derived stem cells(ADSC)to differentiate into Schwann-like cells(iSC).Methods From March,2017 to October,2018,ADSCs were isolated from inguinal adipose tissue of healthy adult female SD rats.Flow cytometry was performed to detect ADSC positive markers CD29,CD90 and negative marker CD45.iSC induction medium was used to culture ADSC.S-100 and GFAP were detected by immunofluorescence staining to confirm that ADSC had differentiated into iSC.Morphological changes of cells were observed by inverted microscope on day 1st,4th,7th,10th,13rd,16th and 19th after induction.MTS assay was used to evaluate cell proliferation ability.Tunel staining was applied to assess cell apoptosis.Results Both S100 and GFAP were expressed in iSC.On day 7th,the cell proliferation rate was significantly slower than that before induction(A value was 0.330±0.020 vs.0.400±0.004,P<0.05).It was negatively correlated with induction time.On day 19th,the proliferation rate of iSC was lower than 50%of the proliferation rate before induction(A value was 0.016±0.003 vs.0.400±0.004,P<0.05).Apoptosis of iSC was more obvious than ADSC at the same time point.Conclusion The proliferation ability of ADSC-induced iSC is optimal within 7 days after induction.

关 键 词：脂肪干细胞 类许旺细胞 许旺细胞 诱导 细胞增殖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]