新生仔猪口腔黏膜上皮细胞的体外培养方法

doi:10.11843/j.issn.0366-6964.2013.09.016

Abstract

Abstract:

The aim of this study was to establish the culture method of piglet oral mucosal epithelial cells in vitro. Piglet buccal cells were cultured and subcultured with the direct explant technique, trypsin digestion, combined method of DispaseⅡand trypsin digestion, and DispaseⅡ combined with explant technique. Morphological characteristics were observed under the light microscope and transmission electron microscope (TEM). For second cultures, the expression of cytokeratin was detected by indirect immunofluorescence, cell growth curve was measured by MTT method，cell cycle distribution was evaluated by flow cytometry. The results showed that in the same culture medium the explant technique and enzymatic method were not successful, the cells obtained by the method of combination of dispaseⅡand trypsin digestion were difficult to attach and grow, but the cells obtained by the combination of DispaseⅡ and explant technique proliferated quickly, and were of high purity, which can be passaged continuously till to the 10-11 generations, and the cells were identified with the characteristics of oral mucosal epithelial cells. The culture method of piglet oral mucosal epithelial cells in vitro has been established successfully, which may provide ideal research material for study the immortalization of oral mucosal epithelial cells, swine foot-and-mouth disease (FMD) pathogenesis and its vaccine.

CLC Number:

Q813.1

MA Rui-li, ZHANG Yan-ming, CUI Hong-jie, GUO Kang-kang. The Culturing Method of Neonatal Piglet Oral Mucosal Epithelial Cells in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(9): 1445-1453.

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The Culturing Method of Neonatal Piglet Oral Mucosal Epithelial Cells in vitro

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