Raft培养可诱导人正常上皮细胞分化

人的正常上皮细胞在一般的体外培养情况下很难分化成类似于机体的上皮样结构。为了得到上皮结构进行相关研究，近年来，国外建立了Raft（船式）培养方法。本文在长期从事细胞培养的工作中，摸索出了适合国内一般实验室条件的Raft细胞培养操作方案，模拟体内人皮肤结构的分化层次，以纤维细胞与胶原作为混合基质（作为上皮细胞的滋养物），同时加入一些生长因子、激素和必需的蛋白等成分。将纤维细胞、胶原和添加成分做为基层过夜培养，次日将培养好的上皮细胞种到上面，过夜培养后即为“skin equivalent”（皮肤等价物），将“皮肤等价物”移到一种网状金属架上以使细胞在气－液状态下培养，在这种条件下，培养中的上皮细胞与体内上皮细胞自然生长的状态、环境非常类似，在体外导致上皮细胞分化成清晰可见的层次。此项技术在临床治疗烧伤以及其它皮肤损伤的疾病方面有潜在的应用价值，也为研究皮肤的正常生理功能以及癌症的发生、发展以及治疗提供了一种模型。

Differentiation results from Raft culture of human epithelium cells

It is very difficult for normal human epithelium cells to differentiate into epitheloid structure in vitro similar to organization in vivo.In recent years,aim to obtain differentiated epithelium structure,Raft culture method has been set up in abroad.Based on the long time cell culture experience,we have established culture protocol sutiable for laboratory use in China.First,collagen and fibroblast were mixed as matrix for the nutritive basal layer,some growth factors,hormones and essential proteins were added and cultured overnight.Second,the epithelium cells were inoculated on the nutritive layer and cotinuously cultured in the following overnight,in the mean time,the equivalent skin were formed.Third,the kin were transferred onto a kind of metal frame so that the epithelium cells could be grown in a disired gas liquid state.The growing condition was similar to that in vivo,by which the epitheliumm cells were led to differentiate and the differentiated graduations were distinctly observed.This protocol has potential values in clinical application such as burn therapy and skin lesion therapy,and it also provides a model in research work as physiological function of human skin and occurrence,progress and therapy for cancer diseases.