人二倍体细胞株(HDCSs),拥有相同的染色体组,已知不含所有已知的不定因子,在开发人类疫苗方面非常有用。然而,获得能够满足疫苗大规模生产要求的合格HDCS是极其困难的。我们开发了一种新的HDCS,Walvax-2,我们从3个月大胎儿的肺组织中获得。我们建立了小学,主和工作细胞库成功地从重建的冷冻细胞。在Walvax-2和MRC-5细胞的同时繁殖过程中的观察显示,在生长速率方面存在差异,细胞活力和病毒敏感性。具体来说,Walvax-2细胞比MRC-5细胞复制更快,Walvax-2细胞在48小时内达到与MRC-5细胞在72小时内达到的相同程度的汇合。此外,Walvax-2细胞获得58代细胞倍增,而MRC-5在此期间达到48代。我们还评估了这些细胞对狂犬病的易感性,甲型肝炎,和水痘病毒.病毒滴度分析显示Walvax-2细胞在培养这些病毒方面等于或优于MRC-5细胞。此外,为了表征Walvax-2细胞库,包括细胞识别在内的一系列测试,染色体特征,致瘤性,以及微生物试剂存在的测试,外源性病毒,和逆转录病毒,是根据标准的国际协议进行的。总之,这项研究的结果表明,Walvax-2细胞库是一种有前途的细胞基质,可用于制造HDCV。
Human diploid cell strains (HDCSs), possessing identical chromosome sets known to be free of all known adventitious agents, are of great use in developing human vaccines. However it is extremely difficult to obtain qualified HDCSs that can satisfy the requirements for the mass production of vaccines. We have developed a new HDCS, Walvax-2, which we derived from the lung tissue of a 3-month-old fetus. We established primary, master and working cell banks successfully from reconstituted frozen cells. Observations during the concurrent propagation of Walvax-2 and MRC-5 cells revealed differences in terms of growth rate, cell viability and viral sensitivities. Specifically, Walvax-2 cells replicated more rapidly than MRC-5 cells, with Walvax-2 cells attaining the same degree of confluence in 48 hours as was reached by MRC-5 cells in 72 hours. Moreover, Walvax-2 cells attained 58 passages of cell doublings whereas MRC-5 reached 48 passages during this period. We also assessed the susceptibility of these cells to rabies, hepatitis A, and Varicella viruses. Analysis of virus titers showed the Walvax-2 cells to be equal or superior to MRC-5 cells for cultivating these viruses. Furthermore, in order to characterize the Walvax-2 cell banks, a series of tests including cell identification, chromosomal characterization, tumorigenicity, as well as tests for the presence of microbial agents, exogenous viruses, and retroviruses, were conducted according to standard international protocols. In conclusion, results from this study show that Walvax-2 cell banks are a promising cell substrate and could potentially be used for the manufacturing of HDCVs.