银狐垂体细胞系构建研究

分别采用组织块法和消化法2种接种方法进行银狐垂体细胞系构建研究。在消化法中使用不同浓度的胰蛋白酶来检验效果,同时在原代培养期使用反复差速贴壁法纯化,传代培养期结合使用反复差速贴壁以及两步消化法进行处理。结果表明：组织块法接种后成纤维细胞生长明显,原代培养后期成纤维细胞已为主要细胞;降低胰蛋白酶浓度能降低对细胞损伤,但消化时间会增长;采用消化法接种在抑制成纤维细胞生长方面效果好于组织块法。纯化效果上,结合反复差速贴壁法与两步消化法处理后获得了较好的效果,在原代培养时期能较好去除成纤维细胞并通过持续使用该方法可在传二代后获得较高纯度的细胞,纯度能达到80%以上,能够建立细胞系。

Establishment of Pituitary Cell Lines of Silver Fox

The pituitary cell lines were tested to establish by tissue explant and digestion explant methods.In the digestion explant method,different concentrations of trypsin were used to test the effect of digestion;in the primary culture,cells were purified repeatedly by differential adhesion method,while in subculture,differential adhesion method and two-step digestion method were used for processing.Fibroblasts grew obviously after inoculation with tissue explant method and became the cell in majority in the late stage of the primary culture;reduction of the concentration of trypsin decreased the damage of cells,but increased the time of digestion;the digestion explant method gave a better result in restricting the growth of fibroblasts than the tissue explant method;combination of the repeated differential adhesion method and the two-step digestion method resulted in better purification;removal of fibroblasts in the primary cell culture and continuous use of the method gave the result of hgher purity of pituitary cells after the second generation,amounting to 80%and cell line were established successfully.