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兔成纤维细胞的分离与体外培养
寇全安1, 张 涌1, 李裕强1
西北农业大学 动物科学与动物医学学院
摘要:
用Ⅰ型、Ⅳ型胶原酶消化分离兔皮肤细胞,Ⅰ型胶原酶消化皮肤块获得的细胞数显著高于Ⅳ型胶原酶。用组织块直接培养法,也可以得到皮肤的原代培养细胞。2 .5 g/L 胰酶消化胎儿组织,可获得足够原代培养的细胞量。结合使用酶消化法和反复贴壁法,经2 ~3代后,可获得纯化的成纤维细胞。分离纯化的胎儿和皮肤成纤维细胞的生长曲线都正常且相似。胎儿和皮肤成纤维细胞可分别传至第13代与第11代,传代后染色体数目不变。
关键词:  皮肤,胎儿,成纤维细胞,分离,体外培养,兔
DOI:
分类号:S814.6
基金项目:农业部生物技术重点项目(95牧-01-04-01)
Dissociation and in vitro Culture of Rabbit Fibroblasts
Abstract:
Rabbit skin tissue digested by collagenase gave a higher yield of cells than by collagenase .The primary culture cells of rabbit skin tissue could be obtained by explant culture too.Rabbit fetal tissue digested by 2.5 g/L trypsin provided a high yield of viable proliferating fibroblast for primary culture.Homogeneous fibroblasts could be obtained after the mixture of fibroblasts and epithelial cells were treated by the combined use of typsin digestion and repeated attachment method for two or three subcultures.The growth curves of fetal and skin fibroblasts were normal and similar.Rabbit fetal and skin fibroblasts could subculture to 13th and 11th generation respectively and still retained the normal chromosome content.
Key words:  skin,fetus,fibroblast,dissociation,in vitro culture,rabbit,