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The medium was half replaced every other day and cells were passaged with Accutase (Sigma) every 7 days and used for subsequent experiments after 24 passages

Various metabolic events caused by these microorganisms form different types and amounts of metabolites throughout the process

The only stipulation is that these gel/ice packs must be frozen solid

[15] In hepatocytes, if all glucose currently stored as glycogen were instead present in its free form, the intracellular glucose concentration would rise to about 0.4 M, compared to the actual polysaccharide concentration of roughly 0.04 M

Nature 451 , 330334 (2008)