Abstract：
Primary cell cultures derived from human embryo lung play a crucial role in virology by aiding virus propagation and vaccine development. These cultures exhibit a notable ability to undergo multiple subcultures, often reaching up to 70 passages. However, finding alternative primary cell cultures with similar longevity and usefulness is challenging. In this study, we introduce a novel primary culture cells derived from equine embryo brain (FEB), which cells exhibited remarkable long-term cultivation potential. The FEB was established and maintained using Sumitomo Nerve-Cell Culture System Comparison studies were conducted with fetal equine kidney cell line (FEK-Tc13) to assess growth rates and subculture longevity. Immunological characterization was performed using neuronal markers to confirm the neural nature of FEB cells. Viral growth assessments were conducted using equine herpesviruses (EHV-1 and EHV-4) to evaluate infectivity and cytopathic effects in FEB cells. PCR analysis and real-time PCR assays were employed to detect viral genomic DNA and transcription activity of EHVs in infected FEB cells. FEB cells demonstrated faster growth rates compared to fetal equine kidney cell line (FEK-Tc13 cells) and exhibited sustained subculture capability exceeding 50 passages. Immunostaining confirmed the glial identity of FEB cells. Both equine herpesviruses 1 and 4 EHV-1 and EHV-4 viruses efficiently replicated in FEB cells, resulting in clear cytopathic effects. PCR analysis detected genomic DNA of EHVs in infected FEB cells, indicating successful viral infection. The establishment of FEB cells with extended subculture capability highlights their potential utility as a model system for studying neural cell biology and viral infections.
摘要：
源自人胚肺的原代细胞培养物通过帮助病毒繁殖和疫苗开发在病毒学中起着至关重要的作用。这些培养物表现出明显的经历多种亚培养的能力，经常达到70个通道。然而,寻找具有相似寿命和实用性的替代原代细胞培养物是具有挑战性的。在这项研究中,我们介绍了一种来自马胚胎脑（FEB）的新型原代培养细胞，这些细胞表现出显著的长期培养潜力。使用住友神经细胞培养系统建立并维持FEB。使用胎马肾细胞系（FEK-Tc13）进行比较研究，以评估生长速率和继代培养寿命。使用神经元标记进行免疫学表征以确认FEB细胞的神经性质。使用马疱疹病毒（EHV-1和EHV-4）进行病毒生长评估，以评估FEB细胞中的感染性和致细胞病变作用。采用PCR分析和实时PCR测定来检测感染的FEB细胞中病毒基因组DNA和EHV的转录活性。与胎马肾细胞系(FEK-Tc13细胞)相比,FEB细胞表现出更快的生长速率,并且表现出超过50代的持续继代培养能力。免疫染色证实了FEB细胞的神经胶质身份。马疱疹病毒1和4EHV-1和EHV-4病毒均在FEB细胞中有效复制，导致明显的细胞病变效应。PCR分析检测到感染的FEB细胞中EHV的基因组DNA，表明成功的病毒感染。具有扩展继代培养能力的FEB细胞的建立突出了其作为研究神经细胞生物学和病毒感染的模型系统的潜在实用性。
