Basic—Liver, Pancreas, and Biliary TractEnrichment and Clonal Culture of Progenitor Cells During Mouse Postnatal Liver Development in Mice
Section snippets
Cell Preparation From Postnatal Livers
Hepatocytes and nonparenchymal cells were isolated from postnatal livers following a 2-step collagenase digestion.26 Perfused liver tissues were subsequently dissociated with 0.05% collagenase solution in 10 minutes at 37°C. The mature hepatocyte fraction was separated from nonparenchymal cells by several episodes of low-speed centrifugation (50g for 1 minute). Nonparenchymal cells were collected by centrifugation (1200 rpm for 5 minutes). Dead cell debris was removed by centrifugation in 25%
Progenitor Cells Derived From CD45−Ter119−c-Kit−Sca1− Population of Normal Postnatal Livers
Treatment with 3,5-diethoxycarbonyl-1,4-dihydrocollidine induces proliferation of oval cells, candidate bipotent progenitor cells, in mouse postnatal livers (“injured livers”).31, 32, 33 The phenotypes of stem/progenitor cells in normal (naive, noninjured) livers remain unknown. To identify the specific surface molecules of PLSCs, we used 2- to 3-week-old mice because preliminary experiments revealed that cells derived from young mice were highly able to form colonies (data not shown).
Discussion
In this study, we established a new primary culture system for clone-sorted cells derived from normal postnatal mouse livers. As shown in Figure 7, we speculate that stem/progenitor cell fraction phenotypes change during liver ontogeny. CD13 and CD133 were common to fetal hepatoblasts and PLSCs, whereas CD49f was detected only on PLSCs. Fetal CD45−Ter119−c-Kit−CD13+CD49f−/lowCD133+ cells and postnatal CD45−Ter119−c-Kit−Sca1−CD13+CD49f+CD133+ cells have similar capacities to form large colonies
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Conflicts of interest The authors disclose no conflicts.
Funding Supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology in Japan, Grants-in-Aid from the Japan KAMPO Medicine Manufacturers Association, and the Kampou Science Foundation.