Detrimental effect of zwitterionic buffers on lysosomal homeostasis in cell lines and iPSC-derived neurons

Good's buffers are commonly used for cell culture and, although developed to have minimal to no biological impact, they cause alterations in cellular processes such as autophagy and lysosomal enzyme activity. Using Chinese hamster ovary cells and induced pluripotent stem cell-derived neurons, this study explores the effect of zwitterionic buffers, specifically HEPES, on lysosomal volume and Ca²⁺ levels. Certain zwitterionic buffers lead to lysosomal expansion and reduced lysosomal Ca²⁺. Care should be taken when selecting buffers for growth media to avoid detrimental impacts on lysosomal function.