【摘要】 目的探索建立少量成年自体皮肤角质形成细胞体外无血清培养体系,为自体组织工程皮肤的构建与移植奠定物质基础。方法:无菌条件下,取2cm×2cm 兔耳皮肤组织块,Dispase 消化,分离真表皮,表皮以胰蛋白酶+EDTA 消化获得角质形成细胞,以含钙和不含钙的角质形成细胞培养液（K-SFM）,按不同细胞密度接种于24孔板,观察细胞生长状况;免疫组化鉴定,MTT 法测定不同血清浓度对角质形成细胞增殖分化的影响。结果:在适当的 Ca²⁺浓度下,少量自体角质形成细胞能够在无血清培养液 k-SFM 中培养扩增,最多可传4～6代。添加血清后可明显加速细胞分化。结论:无血清培养体系适用于少量成年自体角质形成细胞的体外连续培养扩增,培养的细胞可用于自体组织工程皮肤的构建。更多还原

【Abstract】 Objective To explore the culture system of autologous keratiocytes of adult skin in serum free media in vitro,which is the base of autografts of tissue engineering skin.Methods A small piece of skin biopsy about 2cm to 2cm,was got form rabbit ear,digested with dispase,and epidermis was seperated.Keratinocytes were isolated,and cultured in different cell dinsities in the media of keratinocytes were isolated,and cultured in different cell densities in the media of ker- atinocyte—SFM with calcium and without calcium in vitro.The cultured keratinocytes were ob- served,and identificated with lmmunohistochemical test.The effect of K—SFM supplemented dif- ferent contents of new—born bovine serum on keratinocytes was then detected with MTT test. Results The results showed that a small quantities of autologous keratinocytes could growth and proliferated well in K—SFM with suitable content of calcium,and differentiated rapidly under the condition of media supplemented serum.Conclusion It suggested that the culture system of serum free media could be used in proliferation of limited autologous keratinocytes in large scale in vit- ro,which can supplied sufficient cells or tissue engineering.更多还原