Characteristics and viral propagation properties of a new human diploid cell line, Walvax-2, and its suitability as a candidate cell substrate for vaccine production.
Characteristics and viral propagation properties of a new human diploid cell line, Walvax-2, and its suitability as a candidate cell substrate for vaccine production.
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DOI：
10.1080/21645515.2015.1009811
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发表时间：
2015
影响因子：
4.8
通讯作者：
Zheng CY
中科院分区：
文献类型：
--
作者：
Ma B;He LF;Zhang YL;Chen M;Wang LL;Yang HW;Yan T;Sun MX;Zheng CY
人二倍体细胞株（HDCS）具有相同的染色体组，已知不含所有已知的外源因子，在开发人疫苗中有很大的用途。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。我们已经开发了一种新的HDCS，Walvax-2，它来自于3个月大胎儿的肺组织。我们成功地建立了原代细胞库、主细胞库和工作细胞库。在Walvax-2和MRC-5细胞的同时繁殖期间的观察揭示了在生长速率、细胞活力和病毒敏感性方面的差异。具体而言，Walvax-2细胞比MRC-5细胞复制得更快，Walvax-2细胞在48小时内达到与MRC-5细胞在72小时内达到的汇合程度相同的汇合程度。此外，Walvax-2细胞达到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、甲型肝炎和水痘病毒的易感性。病毒滴度分析表明，Walvax-2细胞培养这些病毒的能力等于或上级MRC-5细胞。此外，为了表征Walvax-2细胞库，根据标准国际方案进行了一系列试验，包括细胞鉴定、染色体表征、致瘤性以及是否存在微生物因子、外源性病毒和逆转录病毒的试验。总之，本研究的结果表明，Walvax-2细胞库是一种有前途的细胞基质，可用于生产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.