脂肪干细胞向表皮细胞表型转分化的研究  

Identification and cell phenotype transdifferentiation of adipose-derived stem cells

作　　者：董晓红[1,2] 雷永红[1,3] 付小兵[1] 王文礼[1,2] 孙同柱[1] 李海红[1] 梁晶冰[1] 

机构地区：[1]北京解放军总医院三零四临床部全军创伤修复研究室,100037 [2]内蒙古医学院 [3]第四军医大学西京医院整形外科

出　　处：《中华整形外科杂志》2007年第2期151-153,共3页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金资助项目（30230370）

摘　　要：目的 探讨脂肪干细胞（adipose-derived stem cells，ADSCs）跨胚层向表皮细胞表型转分化的可能性。方法 取大鼠脂肪组织，用酶消化法分离、培养ADSCs，用免疫细胞化学法、流式细胞仪法鉴定ADSCs。实验分为皮肤匀浆条件培养基诱导7d组；皮肤匀浆条件培养基加入EGF（20ng/ml）诱导7d组；皮肤匀浆条件培养基诱导4d，撤条件培养基，改10％FBS／DMEM培养3d组；10％FBS／DMEM为对照组。诱导后经流式细胞仪检测CK19、CK10的表达。结果 ①免疫细胞化学分析表明ADSCs表达CD49d。而不表达CD106、CD34、CK19、CK10；流式细胞仪检测CD49d、CD44的阳性率分别为94．32％、90．38％。②各诱导实验组细胞CK19阳性率分别为45．32％、26．58％、23．37％，CK10阳性率分别为43．56％、25．54％、18．20％，较对照组CK19阳性率18．53％、CK10阳性率2．46％，明显增高（P〈0．01），差异有统计学意义。结论 皮肤匀浆液可以诱导ADSCs向表皮细胞表型转分化。Objective To investigate the transdifferentiation of the ADSCs to epidermal cells. Methods ADSCs were isolated and cultured from rat adipose tissue by digestion of enzyme. ADSCs was identified by immunocytochemistry and flow cytometry. ADSCs were divided into four groups in order to induce： the condition medium（containing 30% superior of homogenating rat skin in 10% FBS/DMEM） group, 7 days; 10% FBS/ DMEM with EGF（20 ng/ml） group, 7 days; the condition medium for 4 clays and then 10% FBS/DEME instead of the condition medium for 3 clays group; 10% FBS/DMEM for 7 days group（ control group）. Cytokeration 19 and cytokeratin 10 expressions in ADSCs were detected by flow cytometry. Results （i） The results of immunocytochemistry showed that ADSCs were positive for CD49d and negative for CD106,CD34,CD19,CD10. The results of flow cytometry showed ADSCs were positive for cIMgd and CD44. Q The CK19 expression of ADSCs was 45.32% in the condition medium group, 26.58% in the condition medium with EGF group, 23.37% in te condition medium for 4 clays and then 10% FBS/DMEM instead of the condition medium for 3 clays gropu and 18.53% in control group, P 〈 0.01. The CK10 expression of ADSCs was 43.56% in the condition medium group, 25.54% in the condition medium with EGF group, 18.20% in the condition medium for4 days and then 10% FBS/DMEM instead of the condition medium for 3 days group and 2.46% in control group, P 〈 0.01. Conclusions The superior of homogenating rat skin can induce CK19 and CK10 expressing in ADSCs, and thereby demonstrating ADSCs can differentiate to epidermal cell phenotype in vitro.

关 键 词：脂肪干细胞 细胞培养 转分化 

分 类 号：R686[医药卫生—骨科学]