以表皮干细胞与hacat细胞为种子细胞构建组织-第三军医大学学报.docVIP

PAGE  PAGE 5 以表皮干细胞与HaCaT细胞为种子细胞构建组织工程皮肤的比较研究 甘 露，曹 川，李世荣△，李建福，毋巨龙（第三军医大学西南医院整形外科，重庆 400038） 摘 要： 目的 比较人表皮干细胞（epidermal stem cells, ESCs）与人永生化角质细胞HaCaT作为种子细胞构建组织工程皮肤的差异。方法 Ⅳ型胶原快速粘附法分选表皮干细胞，无血清培养基DK-SFM培养，取第2代备用；应用DK-SFM培养HaCaT细胞，取对数生长期的细胞备用；利用组织工程技术构建真皮等价物（dermal equivalent）；将上述备用的两种种子细胞分别接种在真皮替代物的表面，构建组织工程皮肤，先后进行浸没培养和气-液面器官型培养，从形态学、物理特性、HE染色等方面观察比较培养物的差异。结果 以ESCs构建的组织工程皮肤收缩快，分层分化近似于正常皮肤，具有较好的张力和韧性。以HaCaT细胞构建的组织工???皮肤分层少，生长慢，张力韧性较差。两种皮肤的直径都随培养时间的增加而缩短，且在3～７ d各检测时相点间有显著性差异（P0.05）。结论 ESCs比永生化的HaCaT细胞更适合作为种子细胞构建组织工程皮肤。 关键词：表皮干细胞；HaCaT细胞；组织工程皮肤 中图法分类号： R318.19；R329.28 文献标识码：A Comparison of epidermal stem cells and HaCaT Cells as seed cells to construct tissue engineered skin GAN Lu1, CAO Chuan2, LI Shi-rong2, LI Jian-fu2, WU Ju-long2（1Out-patient clinic of Chengdu Military Region Air Force Logistics 2Department of Plastic and Aesthetic surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038,China） Abstract：Objective Comparison of human epidermal stem cells and human immortalized keratinocytes HaCaT as seed cells to construct tissue engineered skin differences. Methods Human Epidermal stem cells（ESCs）were selected by rapid attachment to collagen Ⅳ, feed with DK-SFM. Passages 2 were ready to seed. Hacat cells were also feed with DK-SFM. ESCs and Hacat cells were respectively seeded on collagen matrices populated with increasing numbers of fibroblasts and cultured for 1 weeks at the air–liquid interface. From the morphology, physical properties, HE staining and so on observation and comparison of the differences in culture. Results ESCs in constructing a contraction of the tissue-engineered skin faster，stratification is similar to normal skin, has good tension and toughness. HaCaT cells in tissue -engineered skin constructed less laminated, slow growth, low tension and toughness. The diameter of two types of skin along with the increasing of culture time shortened, and in 3 ~ 7 days, the detection time points there was significa