口腔黏膜上皮细胞和成纤维细胞的复合培养

目的 为癌前病变临床病理研究建立一种简便、迅速和有效的口腔黏膜上皮细胞和成纤维细胞复合培养的方法，实现体外模拟组织工程化口腔黏膜的发生发展。方法 用DispaseⅡ分离上皮和皮下组织，用KGM培养口腔黏膜上皮细胞。用细胞培养法和组织块培养法获取验用的口腔黏膜上皮细胞和成纤维细胞，并采用复合培养法对两种细胞进行共同培养。结果 用DispaseⅡ可成分地分离上皮和皮下组织。KGM可明显促进口腔黏膜上皮细胞的分裂繁殖。复合培养的HE染色切片显示薄层的结缔组织之上有方形的基底细胞、颗粒细胞和角化层。结论 KGM可明显促进口腔黏膜上皮细胞的分裂和成熟。采用组织培养法获取原代成纤维细胞是适合口腔黏膜取材等特点的有效方法。气液相培养的口腔黏膜下结缔组织可促进上皮细胞的分层和分化。

Coculture of oral epithelial cells and fibroblasts

Objective To establish a simple, reliable, rapid and efficient oral epithelial cell and fibroblast coculture technique in vitro for the purpose of the premalignant physiopathologic study in clinic. Methods Dispase II was used to separate epithelium from the underlying fibrous connective tissue. The oral epithelial cells were grown in keratinocyte growth medium( KGM) . The primary fibroblasts were obtained by tissue culture and the oral epithelial cells were obtained by cell culture. Then the tissue speciment of oral epithelial and fibroblast were obtained by coculture of two types of cells. Results Dispase II can successfully separate epithelium from the underlying fibrous connective tissue. KGM can enhance oral mucosa epitheeium growth and propagating. Section of 12 days coculture of keratinocytes and fibroblasts showed the epidermis in vitro consists of cuboidal cells. a squamous layer and an upper horny. Layer on the dermis consists of a few fibroblasts. Conclusion The results mentioned shove indicate that KGM can enhcance the oral mucosa growth an propagating. It is an effective method to obtain fibroblasts by tissue culture. Coculture of keratinocytes and fibroblasts in liguid/air condition can improve epidermal stratify and differentiation.