A serum-free medium suitable for maintaining cell morphology and liver-specific function in induced human hepatocytes
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hiHep is a new type of hepatocyte-like cell that is predicted to be a potential unlimited source of hepatocytes for a bioartificial liver. However, hiHep cannot currently be used in clinical settings because serum must be added during the culture process. Thus, a defined medium is required. Because serum is complex, an efficient statistical approach based on the Plackett–Burman design was used. In this manner, an original medium and several significant cell growth factors were identified. These factors include insulin, VH, and VE, and the original medium was optimized based on these significant factors. Additionally, hiHep liver-specific functions and metabolism in the optimized serum-free medium were measured. Results showed that hiHep functions, such as glycogen storage, albumin secretion, and urea production, were well maintained in our optimized serum-free medium. In summary, we created a chemically defined, serum-free medium in which cell growth, proliferation, metabolism, and function were well maintained. This medium has the potential to support the clinical use of hiHep.
KeywordshiHep Serum-free Medium Bioartificial liver
We thank Lijian Hui’s lab for providing hiHep and the cell culturing protocol. We are also grateful to Xuan Ni from Guoyu Pan’s lab for performing CYP activity quantification. This research was supported by the National Natural Science Foundation of China (Grant Nos. 31170951 and 81671841), the Natural Science Foundation of Shanghai (Grant No.16ZR1408700) and Basic Research Key Program Project of Commission of Science and Technology of Shanghai (16JC1400203).
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