中文摘要：

为了研究猪胎儿成纤维细胞和耳皮肤成纤维细胞的分离培养体系，根据取样组织的不同，筛选出最佳的培养方法，试验以猪胎儿和耳皮肤为试验材料，用4种不同的培养分离方法，即胰酶热消化法、胰酶冷热结合消化法、胰酶室温消化法和组织块培养法，分离培养猪胎儿成纤维细胞和耳皮肤成纤维细胞，对比培养效果，筛选出最佳的培养方法。结果表明：胰酶室温消化法分离的猪胎儿成纤维细胞存活率显著高于胰酶热消化法和胰酶冷热结合消化法（P〈0．05），细胞贴壁效果好，且操作简单；猪耳皮肤成纤维细胞原代培养中，胰酶热消化法和胰酶室温消化法分离的细胞数量少，组织块培养法所需的组织量少，且较胰酶冷热结合消化法操作简单。试验中培养的细胞呈典型的成纤维细胞形态，生长曲线呈“S”型，经冻存复苏后生长状态良好，说明胰酶室温消化法是猪胎儿成纤维细胞较好的培养方法，胰酶热消化法和胰酶室温消化法是猪耳皮肤成纤维细胞原代培养的理想培养方法。

英文摘要：

To study the isolated culture system of porcine fetal fibroblasts and ear skin fibroblasts for selecting the best culture method according to the samples from different tissues, porcine fetal and ear skin were used as experimental material, the porcine fetal fibroblasts and ear skin fibroblasts were isolated and cultured by four different methods including the methods for trypsin hot digestion, trypsin digestion of hot and cold combination, trypsin digestion at room temperature and tissue culture. The effects of the four methods were compared to find a better one. The re- suits showed that the survival rate of porcine fetal fibroblasts isolated by the method for trypsin digestion at room temperature, was significantly higher than those by the methods for trypsin hot digestion and trypsin digestion of hot and cold combination （ P 〈 O. 05 ）. The method for trypsin digestion at room temperature was simple to operate, and the effect of cell adhesion was better. In primary cultured ear skin fibroblasts, a small number of cells were isolated by the methods for trypsin hot digestion and trypsin digestion at room temperature. The method for tissue culture needed a small number of tissues, and was easier to operate than the method for trypsin digestion of hot and cold combination. The cultured cells were in typical fibroblast morphology, the growth curve was sigmoidal, and the cells grew well after freezing and thawing. The results indicate that the method for trypsin digestion at room temperature is a good culture method is for porcine fetal fibroblast, and the methods for trypsin hot digestion and the trypsin digestion at room temperature are the ideal culture methods for primary cultured porcine ear skin fibroblasts.