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  

Comparison of epidermal stem cells and HaCaT cells as seed cells to construct tissue engineered skin

作　　者：甘露[1] 曹川[1] 李世荣[1] 李建福[1] 毋巨龙[1] 

机构地区：[1]第三军医大学西南医院整形外科,重庆400038

出　　处：《第三军医大学学报》2009年第20期1977-1980,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30600652)~~

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型胶原快速黏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(dermalequivalent);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细胞构建的组织工程皮肤分层少,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3～7d各检测时相点间有显著性差异(P<0.05)。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。Objective To explore the differences of human epidermal stem cells （ESCs） and human immortalized keratinocytes HaCaT as seed cells to construct tissue engineered skin. Methods Human ESCs were selected by rapid attachment to collagen Ⅳ, and fed with DK-SFM. The cells at passages 2 were ready to seed. Hacat cells were also fed with DK-SFM. ESCs and Hacat cells were respectively seeded on collagen matrices populated with increasing numbers of fibroblasts and cultured for 1 week at the air liquid interface. Morphology, physical properties observation, and HE staining were carried out to compare the differences of the 2 cells. Results ESCs were faster in constructing a contraction of the tissue-engineered skin, and had good tension and toughness and similar stratification as normal skin. HaCaT cells were less laminated, slow growth, and had low tension and toughness in tissue-engineered skin construction. The diameter of 2 types of constructed skin samples was shortened along with the increasing of culture time, and had significant difference in 3 to 7 d. Conclusion Epidermal stem cells are more suitable as seed cells for tissue engineering of skin than the HaCaT immortalized cells.

关 键 词：表皮干细胞 HACAT细胞 组织工程皮肤 

分 类 号：R318[医药卫生—生物医学工程] R322.99[医药卫生—基础医学]