人角质形成细胞复合异种脱细胞真皮基质三维培养的形态学观察  

Morphological observation of human keratinocytes tridimensionally cocultured with xenogenic acellular dermal matrix

作　　者：朱崇涛[1] 彭代智[1] 王丽华[1] 何斌[1] 郑必祥[1] 周新[1] 罗海水[1] 王勇[1] 刘敬[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室,重庆400038

出　　处：《中华烧伤杂志》2009年第4期258-260,共3页Chinese Journal of Burns

基　　金：国家重点基础研究发展计划（2005CB522605）;国家高技术研究发展计划（2006AA02A121）

摘　　要：目的建立组织工程皮肤三维培养方法，了解人永生化KC株（HaCaT）复合异种ADM培养的组织形态学变化。方法采用改良法制备sD大鼠ADM，培养HaCaT。将处于对数生长期的HaCaT按2×10^5个／cm^2密度接种存ADM上浸没培养5d，再转为气液相培养5d。扫描电镜观察ADM及其复合HaCaT培养第1天和第5天的细胞生长情况，HE染色观察ADM及其复合HaCaT培养1、5、10d的组织学变化。结果ADM为白色，外观平整质地柔软，胶原束完整，未见细胞残留。HaCaT接种于ADM培养1d即附着并伸出伪足，5d融合成片，10d增长至3～6层并有向ADM内生长的趋势。结论SD大鼠ADM有利于HaCaT黏附及培养液渗透，可以在10d内获得复层组织工程皮肤。Objective To establish the tridimensional culture method for tissue-engineered skin to observe the histomorphological change in human immortal KC strain （HacaT） cocultured with xenogenie acel- lular dermal matrix （ADM）. Methods The ADM was prepared from SD rats by a modified method. HaCaTs were cultured in defined KC-serum free medium. HaCaTs in log growth phase were inoculated on ADM at the cell density of 2 × 10^5/em^2. They were submergedly cultured for 5 days and then changed to air- liquid phase culture for another 5 days. ADM and growth of HaCaTs on day 1 and 5 after cocultured with ADM were observed with scanning electron microscope. The histological change in ADM and HaCaTs on day 1, 5, and 10 after cocultured with ADM were examined by HE staining. Results The gross appearance of ADM was white with smooth and soft texture, and intact collagen bundles without cellular residue. HaCaTs adhered and stretched out pseudopodia on the surface of the ADM on day 1 after combined euhure, and a monolayer of cells was formed on day 5, growing into 3 -6 layers of cells on day 10 with a tendency to grow into ADM. Conclusions SD rats ADM is benefit for the adhesion of HaCaTs and the permeation of nutrient solution, from which an engineered multiple-layered human skin can be obtained within 10 days.

关 键 词：真皮 共同培养技术 皮肤 人工 角质形成细胞 形态学 

分 类 号：R318.0[医药卫生—生物医学工程]