摘要
目的探讨脂肪干细胞(ADSCs)经诱导向表皮细胞转分化的可能性。方法(1)切取12例SD大鼠脂肪组织,用酶消化法分离和培养ADSCs,流式细胞仪检测第3代细胞的CD49d、CD44和CD106的表达以鉴定ADSCs;(2)诱导大鼠ADSCs向表皮细胞转分化的实验分组和处理:①含30％大鼠皮肤匀浆液的条件培养基组;②条件培养基中加入EGF(20μg／L)组;③含EGF(20μg／L)的10％FBS DMEM组;④10％FBS DMEM为对照组,各组诱导3 d。(3)诱导后经流式细胞仪检测:①各组ADSCs CK19、CK14的表达。②条件培养基组诱导前、3 d后ADSCs细胞周期。结果流式细胞仪检测CD49d、CD44和CD106阳性率分别为98．32％、90．38％和0．06％;各诱导组细胞CK19阳性率分别为10．44％、8．22％、4．49％,CK14阳性率分别为12．46％、8．19％、4．37％,均较对照组CK19阳性率1．38％、CK14阳性率1．7％升高(P<0．01),差异有统计学意义。条件培养基诱导前ADSCs细胞周期:G1:67．25％、G2:2．46％、S:30．29％,诱导3 d后ADSCs细胞周期: G1:35．04％、G2:0．00％、S:64．96％,细胞分裂增殖速度加快。结论ADSCs可以被诱导向表皮细胞表型转分化,提示ADSCs具有参与皮肤创面修复的潜能。
Objective To investigate the cellular phenotype conversion of adipose derived stem cells （ADSCs） to epidermal cells. Methods （1） ADSCs were isolated and cultured from rat adipose tissue （ n = 12） ,and P3 ADSCs were identified by detecting the expression of CD49d,CIM4 and CD106 by flow cytometry. （2） ADSCs were induced for 3 days by different revulsion, and the expression of cytokeratin 19 and cytokeratin 14 in ADSCs was detected by flow cytometry. Following groups were allocated： the condition medium group, group of the condition medium containing EGF, group of 10% FBS DMEM with EGF and control group. （3） By using flow cytometry,cell cycle in condition medium groups and the control group was analyzed. Resuks （1） Flow cytometry revealed that the positive rate of CD49d, CD44 and CD106 in ADSCs was 98.32% ,90.38% and 0.06% respectively; （2） The positive rate of CK19 was 10.44 % ,8.22 % ,4.49% ,and of CK14 was 12.46 %, 8.19 %, 4.37 % in condition medium group, group of condition medium containing EGF and group of 10 % FBS DMEM with EGF respectively, and all of them were higher than that in control group （1.38 % for CK19 and 1.7 % for CK14 respectively, P 〈 0.01 ） ; （3） The cell cycle was Gl ： 35.04 %, G2 ： 0.00 %, S： 64.96 % at condition medium group after 3 days, and Gl ： 67.25 %, Gz ： 2.46 %, S： 30.29 % at control medium group, suggesting that the skin homogenate could promote the proliferation of ADSCs. Conclusion The homogenate of rat skin can induce the expression of CK19 and CK14 in ADSCs,suggesting ADSCs had potential to differentiate into epidermal cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第12期1536-1538,共3页
Chinese Journal of Experimental Surgery
基金
国家重点基础研究发展规触资助项目(2005CB522603)
国家自然科学基金资助项目(30400172)
全军"十一五"计划资助项目(069116)
关键词
成体干细胞
细胞培养
分化
Adult stem cells
Cell culture
Differentiation
