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一种新的人二倍体细胞系RAZI-HDC的增殖特性及其与MRC-5相比作为呼吸道合胞病毒疫苗生产候选细胞基质的适用性

Propagation Properties of a New Human Diploid Cell Line, RAZI-HDC, and Its Suitability as a Candidate Cell Substrate for Respiratory Syncytial Virus Vaccine Production in Comparison to MRC-5.

作者信息

Abbasi A, Mohammadi A, Alirezaie B, Esna-Ashari F, Sadri N, Salimi V

机构信息

Department of Biotechnology, Faculty of Advanced Sciences and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.

Razi Vaccine and Serum Research Institute (RVSRI) Hessark Karadj Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran.

出版信息

Arch Razi Inst. 2025 Feb 1;80(1):51-59. doi: 10.32592/ARI.2025.80.1.51. eCollection 2025 Feb.

Abstract

Respiratory syncytial virus (RSV) is a common cause of infection of the respiratory tract in infants, older adults, individuals with heart and lung disease, and immunocompromised patients. The disease is responsible for between 100,000 and 200,000 infant deaths on an annual basis. RSV vaccine production platforms have been developed. In this study, a local diploid cell line, RAZI-HDC, derived from human fetal lung cells, was utilized for RSV virus propagation with the objective of studying live-attenuated vaccine, and was compared to the MRC-5 cell line. The total cells per 25-cm flask were 44.0 ± 2.6 *105 and 41.66 ± 2.08 *105 for MRC-5 and RAZI-HDC, respectively. The maximum cell-specific growth rate of RAZI-HDC was 316.66 ± 20.81, while that of MRC-5 was only 340 ± 26.45. The maximum cell division number of RAZI-HDC was 1.24 ± 0.07, in comparison to the MRC-5, with a maximum cell division number of 1.32 ± 0.08. Both cell substrates achieved maximum cell density five days after the initiation of the culture. The complete cytopathic effect of RSV in RAZI-HDC-RAZI-HDC was observed after four days, indicating the sensitivity of these cells to RSV. The virus productivity in RAZI-HDC cells (2.4685) was not significantly different from that in MRC-5 cells (2.5), as determined by a two-tailed t-test (p=0.78). The results indicated that both cell substrates function similarly in terms of RSV propagation. It is noteworthy that diploid cell lines, such as MRC-5 and RAZI-HDC, are particularly well-suited for vaccine manufacturing. This is primarily due to their human origin and the stability of their karyotype. This is a significant advantage, as it helps ensure the safety of the final vaccine product if these cells are used to make viral vaccines that require virus amplification. The study further assessed the replication capacity of the RAZI-HDC cell line and found it to be equivalent to that of the MRC-5 cell line. Specifically, the maximum virus productivity in RAZI-HDC cells (2.4685 log TCID50/mL) was not significantly different from that in MRC-5 cells (2.5 log TCID50/mL), as determined by statistical analysis. The utilization of a locally developed cell line such as RAZI-HDC has the potential to be more cost-effective in comparison to relying on imported cell substrates.

摘要

呼吸道合胞病毒(RSV)是婴儿、老年人、患有心肺疾病的个体以及免疫功能低下患者呼吸道感染的常见病因。该疾病每年导致10万至20万婴儿死亡。RSV疫苗生产平台已经研发出来。在本研究中,一种源自人胎儿肺细胞的本地二倍体细胞系RAZI-HDC被用于RSV病毒繁殖,目的是研究减毒活疫苗,并与MRC-5细胞系进行比较。对于MRC-5和RAZI-HDC,每25平方厘米培养瓶中的细胞总数分别为44.0±2.6×10⁵和41.66±2.08×10⁵。RAZI-HDC的最大细胞比生长速率为316.66±20.81,而MRC-5的仅为340±26.45。RAZI-HDC的最大细胞分裂数为1.24±0.07,相比之下,MRC-5的最大细胞分裂数为1.32±0.08。两种细胞底物在培养开始后五天达到最大细胞密度。在四天后观察到RSV在RAZI-HDC - RAZI-HDC中出现完全细胞病变效应,表明这些细胞对RSV敏感。通过双尾t检验确定,RAZI-HDC细胞中的病毒生产力(2.4685)与MRC-5细胞中的(2.5)无显著差异(p = 0.78)。结果表明,两种细胞底物在RSV繁殖方面功能相似。值得注意的是,二倍体细胞系,如MRC-5和RAZI-HDC,特别适合疫苗生产。这主要是由于它们源自人类且核型稳定。这是一个显著优势,因为如果使用这些细胞来生产需要病毒扩增的病毒疫苗,有助于确保最终疫苗产品的安全性。该研究进一步评估了RAZI-HDC细胞系的复制能力,发现其与MRC-5细胞系相当。具体而言,通过统计分析确定,RAZI-HDC细胞中的最大病毒生产力(2.4685 log TCID₅₀/mL)与MRC-5细胞中的(2.5 log TCID₅₀/mL)无显著差异。与依赖进口细胞底物相比,使用如RAZI-HDC这样本地开发的细胞系可能更具成本效益。

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