Large scale manufacture of human gallbladder epithelial cells to generate hepatocytes via chemical defined approach for clinical utility
Generation of liver stem cells hold a great promise for treating liver disease or drug screening. We established a chemical defined and xeno-free approach to manufactured large-scale human gallbladder epithelial cells (hGBECS) in GMP condition for clinical utility. The cell products were characterization of growth ability, phenotype, frozen-thaw viability, genetic stability, bio-safety assessments for in vitro tests, and tumorigenicity assessments, acute toxicity experiments for in vivo tests. We then established a protocol to generate functional hepatocytes from hGBECS (hGBECS-dHEPs). The derived hepatocytes exhibited typical liver function: glycogen storage, lipid synthesis, uptake ability, Albumin secretion, urea production and drug metabolism. Also, these cells were applied for tests of liver toxic drugs and anticancer drugs. We further assessed these derived hepatocytes for Cu2+ transportation and alcohol metabolism. Transplantation of these cells in vivo could rescue mice from liver failure. We provide these large-scale, rapid, safe source of functional hepatocytes from hGBECS to serve as a bio-bank for clinical application of treating liver disease.