鸡胚细胞的传代培养和麻疹病毒的增殖

为了建立原代鸡胚细胞的传代培养工艺,探究传代鸡胚细胞对麻疹病毒的敏感性和适应性,本研究将原代鸡胚细胞进行传代培养,分别采用原代鸡胚细胞和传代鸡胚细胞培养麻疹病毒沪-191(Shanghai-191,S-191)株毒种,并对病毒收获液进行滴度检测和基因序列测定。结果显示,原代鸡胚细胞可稳定传代培养至第10代,各代次细胞生长趋势相似;第5代鸡胚细胞染色体检查为正常染色体核型;第8代鸡胚细胞成瘤性检查未见成瘤;采用第3、5代鸡胚细胞制备的麻疹病毒滴度水平高于原代鸡胚细胞,但无显著性差异(n=3,P>0.05),编码病毒核蛋白(nucleoprotein,N)和血凝素蛋白(hemagglutinin,H)的基因序列与S-191株完全一致,未发生变异。本研究证实,原代鸡胚细胞可进行传代培养,各代次鸡胚细胞对麻疹病毒的敏感性不变,产毒水平无显著差异,可用于培养麻疹病毒。

Subculture of chicken embryo cells for propagation of measles virus

To establish the subculture technology of primary chicken embryo cells and explore the sensitivity and adaptability of passage chicken embryo cells to measles virus, the primary chicken embryo cells were subcultured, and the measles virus Shanghai-191 (S-191) strain was cultured in primary chicken embryo cells and subcultured chicken embryo cells. The titer of the virus harvest liquid was detected and the gene sequence was determined. The results showed that the primary chicken embryo cells could be stably subcultured to the 10th generation, and the growth trend of each generation was similar. The chromosome examination of the 5th generation chicken embryo cells showed normal chromosome karyotype, and the tumorigenicity test of the 8th generation chicken embryo cells showed no tumorigenesis. The titers of measles virus prepared from the 3rd and 5th passage of chicken embryo cells were higher than that of the primary chicken embryo cells, but there was no significant difference (n＝3, P＞0.05). The gene sequences of the nucleoprotein(N)and hemagglutinin(H)of the virus were completely consistent with those of S-191 strain, and there was no variation. This study confirms that the primary chicken embryo cells can be subcultured, and the sensitivity of each generation of chicken embryo cells to measles virus remains unchanged. There is no significant difference in the level of measles virus production, which could be used to culture measles virus.