摘要
目的探讨ES源表皮干细胞（ESC）与高分子材料构建人工皮肤Inspectskin Ⅰ的方法，为化妆品皮肤毒性检验替代模型奠定基础。方法采用嵌入式培养法，先将3-羟基丁酸-co-3-羟基戊酸共聚物（PHBV）支架与原代小鼠成纤维细胞共培养5d构建真皮，再于表面放置ESC细胞，气-液界面培养7—10d，形态学和β1整合素、CK10、CK19免疫组化观察。结果成纤维细胞可在真皮支架内粘附，ESC细胞可在支架表面增殖分化形成含基底层、棘层和角化层的复层表皮结构，免疫组化表明基底层以上细胞中表达CK10明显增多，基底层有些细胞仍呈β1整合素和CK19阳性。结论Inspectskin Ⅰ替代皮肤模型是由ESC细胞分化形成的角化复层表皮结构和PHBV真皮结构共同构成的全层皮肤模型。
Objective To investigate the methods of constructing alternative skin Inspectskin Ⅰ by ES-derived epidermal stem cells（ESC） and polymeric materials, and to lay a basis for seek new alternative model for cosmetics skin toxicity assessment. Methods Using the insert culture system, the dermis were cultured with primary mice fibroblast in polyβ-hydroxybutyric acid-co-3- hydroxyvaleric acid]（PHBV） scaffolds for 5 ds, then the ESC cells were placed upside. After air-liquid cultured in vitro for 7 - 10 ds, the morphological and β1 integrin, CK10, CK19 immunohistochemical staining were detected. Results The fibroblast adhered inside the PHBV scaffolds and ESC were differentiated and proliferate into stratified squamous epidermal. Histochemical staining showed that the CK10 expressed ceils were found in all sup-basal, some cells in stratum basal were still expressed β1 integrin and CK19. Conclusion The Inspectskin Ⅰ was the full-thick alternative skin model by ESC differentiating into stratified epidermal and PHBV forming dermal.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2008年第6期428-431,共4页
Journal of Toxicology
基金
国家自然科学基金项目(30670541)
广东省科技计划项目(2005B32401010)
关键词
化妆品
组织工程皮肤
替代模型
Cosmetics
Tissue engineering skin
Alternative model
