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The successful establishment and long-term maintenance of human cell lines require strict control of the culture conditions to ensure phenotypic stability, genomic integrity, and experimental reproducibility. This review provides practical guidelines for culturing established and newly derived cell lines, focusing on three core factors: substrate coating, culture media composition, and environmental consistency. Appropriate coating materials are essential for supporting cell adhesion and lineage-specific function. Culture media must be carefully selected and supplemented to match the metabolic and differentiation needs of the target cell type. Additionally, environmental parameters such as oxygen concentration should be optimized. Consistent passaging protocols and enzymatic dissociation techniques as well as regular authentication are also essential for maintaining cell identity and function. Together, these strategies form the foundation for reliable in vitro cell culture, enabling applications in disease modeling and regenerative medicine.

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Maintaining the Genotype and Phenotype of Human Cell Lines Through Optimization of Culture Conditions, Passaging, and Differentiation Induction

  • 章节

  • 发表日期：

    2025年

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  • Aiki Marushima,
  • Hiroshi Ishikawa

摘要

The successful establishment and long-term maintenance of human cell lines require strict control of the culture conditions to ensure phenotypic stability, genomic integrity, and experimental reproducibility. This review provides practical guidelines for culturing established and newly derived cell lines, focusing on three core factors: substrate coating, culture media composition, and environmental consistency. Appropriate coating materials are essential for supporting cell adhesion and lineage-specific function. Culture media must be carefully selected and supplemented to match the metabolic and differentiation needs of the target cell type. Additionally, environmental parameters such as oxygen concentration should be optimized. Consistent passaging protocols and enzymatic dissociation techniques as well as regular authentication are also essential for maintaining cell identity and function. Together, these strategies form the foundation for reliable in vitro cell culture, enabling applications in disease modeling and regenerative medicine.