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<p>Lentiviral vectors (LVs) have revolutionized gene therapy by enabling stable gene integration into dividing and non-dividing cells, addressing critical challenges in treating genetic disorders. The transition from second to third-generation LVs has increased biosafety by minimizing the risk of replication-competent lentiviruses and expanded their clinical applicability. Despite significant advancements, producing high-titer functional LVs, particularly at an industrial scale, remains a considerable challenge due to the need for enhanced scalability, cost-efficiency, and effectiveness. This Review delves into cutting-edge innovations in LV production, from optimized transient transfection in various cell lines to the development of stable producer cell lines. Stable producer cell lines offer unparalleled scalability but face challenges related to viral protein cytotoxicity. Inducible systems have emerged as pivotal tools for addressing these problems, allowing for precise gene expression and controlled production. Additionally, advancements in bioprocess engineering, ranging from optimized culture conditions, including pH and media composition, to novel bioreactor technologies like structured fixed-bed systems, continue to redefine industrial-scale LV production. These breakthroughs, coupled with the analysis of costs and efficiencies of various methodologies, can further illustrate the potential for large-scale LV production and facilitate widespread therapeutic applications.</p>

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Upstream insights for lentiviral vector production: cell platforms, culture parameters, and titer yields

  • 期刊论文

  • 发表日期：

    2026年3月3日

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  • Pedram Abdollahpour,
  • Alireza Shafizadeh,
  • Ehsan Arefian

摘要

Lentiviral vectors (LVs) have revolutionized gene therapy by enabling stable gene integration into dividing and non-dividing cells, addressing critical challenges in treating genetic disorders. The transition from second to third-generation LVs has increased biosafety by minimizing the risk of replication-competent lentiviruses and expanded their clinical applicability. Despite significant advancements, producing high-titer functional LVs, particularly at an industrial scale, remains a considerable challenge due to the need for enhanced scalability, cost-efficiency, and effectiveness. This Review delves into cutting-edge innovations in LV production, from optimized transient transfection in various cell lines to the development of stable producer cell lines. Stable producer cell lines offer unparalleled scalability but face challenges related to viral protein cytotoxicity. Inducible systems have emerged as pivotal tools for addressing these problems, allowing for precise gene expression and controlled production. Additionally, advancements in bioprocess engineering, ranging from optimized culture conditions, including pH and media composition, to novel bioreactor technologies like structured fixed-bed systems, continue to redefine industrial-scale LV production. These breakthroughs, coupled with the analysis of costs and efficiencies of various methodologies, can further illustrate the potential for large-scale LV production and facilitate widespread therapeutic applications.