Purification of Hepatoblasts from Fetal Mouse Livers by Using a Temperature-Reversible Gelation Polymer and Their Application in Regenerative Medicine
A convenient and efficient purification protocol for fetal mouse hepatoblasts and hepatocytes, which used a thermo-reversible gelation polymer (Mebiol), was established. When dispersed fetal mouse liver cells at E12.5 were cultured within the Mebiol gel, hepatoblasts rapidly aggregated into spheroids, and gave rise to hepatocytes expressing urea cycle enzymes on day 5. By contrast, nonparenchymal cells such as hepatic stellate cells and endothelial cells were dead within the gel. After culture for 3 or 5 days, fetal hepatoblast/hepatocyte aggregates could be easily separated from dead, single nonparenchymal cells by filtration. When E12.5 liver cells were cultured in a Mebiol gel containing basal laminar components, hepatoblasts gave rise to biliary epithelial cells forming cystic structures and expressing no hepatocyte marker. The culture and purification protocol for fetal liver progenitor cells may greatly contribute to regenerative medicine that aims at elucidating their growth and maturation mechanisms, and at developing novel hybrid-type artificial liver models and cell transplantation therapy.
KeywordsHepatic Stellate Cell Lower Critical Solution Temperature Gelation Polymer Nonparenchymal Cell Biliary Epithelial Cell
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