下表列举了进行细胞培养实验时的一些潜在问题，以及可帮助排除故障的可能解决方案。请注意，以下列表仅包括细胞培养中最常见的问题，并且仅提供解决方案指南。为了帮助您更成功地评估结果，我们建议您查阅您使用产品随附的手册和产品信息表，以及已发表的相关文献和书籍。

问题：细胞储存液解冻后没有活细胞

原因
解决方案
细胞储存不当
  • 获取新的细胞储存液并储存于液氮中。 将细胞保存在液氮中，直至解冻
自制冷冻细胞储存液中没有活细胞
  • 按照供应商推荐的密度冻存细胞
  • 使用低传代细胞制作冷冻细胞储存液
  • 严格按照供应商建议的程序冷冻细胞。请注意，本手册推荐的冷冻程序是一般程序，仅供参考。
  • 获取新的细胞储存液
细胞解冻不当
  • 严格按照供应商建议的程序复苏细胞。请注意，本手册推荐的解冻程序是一般程序，仅供参考。
  • 确保快速解冻冷冻细胞，但在接种前使用预热的生长培养基缓慢稀释细胞。
复苏培养基不合适
  • 使用供应商推荐的培养基。确保预热培养基
细胞稀释过度
  • 按照供应商的建议，以高密度接种解冻后的细胞，优化回收率
细胞处理过程不够轻柔
  • 大多数细胞在冷冻和解冻时均受到外力作用。 请勿涡旋、敲击培养瓶使细胞脱落（培养昆虫细胞时除外），或是高速离心细胞。
冷冻培养基中使用的甘油未避光储存（如适用）。
  • 如果未避光储存，甘油会转化成对细胞有毒性的丙烯醛。获取新储备液。

问题：细胞生长缓慢

原因
解决方案
生长培养基不合适
生长培养基中的血清质量不良
  • 使用其他批次的血清。
细胞传代次数过多
  • 使用健康的低传代数细胞。
让细胞生长超过汇合状态
  • 在哺乳动物细胞达到汇合状态前处于对数期时进行传代。
培养物被支原体污染
  • 丢弃细胞、培养基和试剂。获取新的细胞储备液，并将其与新鲜培养基和试剂配合使用。

Gibco 细胞培养基础知识视频

Getting started in 3D - Setting up your lab and reviewing common pitfalls of researchers new to 3D
Part 1: Getting started in 3D: Setting up your lab and reviewing common pitfalls of 3D research Part 2: Monitoring your 3D culture (what's good and bad): How to detect problems and what a successful growth looks like
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