Long-term Extensive Expansion of Mouse Hepatic Stem/Progenitor Cells in a Novel Serum-Free Culture System
Open accessAbstract
Background & aims The liver has high regenerative potential. We attempted to establish a novel culture system for extensive expansion of fetal mouse hepatic stem/progenitor cells and to characterize cultured cells.Methods Hepatic spheroids collected from 6-day floating cultures were cultured on collagen-coated dishes in serum-free conditions in medium containing growth factors. Cultured cells were mainly characterized by immunocytochemistry and flow cytometry or transplanted into adult mice.Results Approximately 400 expanding hepatic spheroids were generated from every 1 x 10(6) fetal liver cells. Subsequently, highly replicative colonies were subcultured with maintaining colony formation on collagen-coated dishes. These colonies consisted of small immature alpha-fetoprotein-positive cells and hepatocytic and cholangiocytic lineage-committed cells. The immature alpha-fetoprotein-positive cells could be expanded in a reproducible manner at least 5 x 10(5)-fold (which involved at least 30 passages over >6 months) without losing differentiation potential. Flow cytometric analysis showed that all cultured cells expressed CD49f, but not CD34, Thy-1, c-kit, or CD45. Nearly 15% of the cells expressed Sca-1, and approximately 5%-20% of the cells were side population cells. Both sorted side population cells and Sca-1-positive cells (especially side population cells) produced a large number of alpha-fetoprotein-positive cells and lineage-committed cells. Expanded cells had bidirectional differentiation potential and improved serum albumin levels in mice with severe liver damage.Conclusions Long-term extensive expansion of transplantable hepatic stem/progenitor cells was reproducibly achieved in a novel serum-free culture system. Moreover, this culture system yielded side population and Sca-1-positive cell populations that included hepatic stem/progenitor cells with differentiation and proliferation properties.
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Using a new passaging method where cells are detached with highly diluted trypsin and lifted and replated as colonies, the viability and phenotypic stability of passaged cdHPCs increase.
“Using this method we found that the viability and phenotypically stability of passaged cdHPCs increased (Supplementary figure 4a).”
From Hepatic progenitor cells of biliary origin with liver repopulation capacity (2015) · cites this paper as [38]
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Self-citation
- Authors
- Tsuchiya, A., Heike, T., Fujino, H., Shiota, M., Umeda, K., Yoshimoto, M., Matsuda, Y., Ichida, T., Aoyagi, Y., & Nakahata, T.
- Affiliations
- Kyoto University (JP)
- Niigata University (JP)
- Juntendo University (JP)
- Corresponding author
- Nakahata, Tatsutoshi
- Venue
- Gastroenterology
- Publisher
- Elsevier BV
- Published
- 2005-06
- Volume
- 128
- Issue
- 7
- Pages
- 2089-2104
- ISSN
- 0016-5085 (ISSN-L)
- OpenAlex
- W1967598939
- Cited by
- 43
- References
- 41
- Citations by year
- 2020: 1 · 2019: 2 · 2018: 3 · 2017: 2 · 2016: 1 · 2015: 2 · 2014: 3 · 2013: 3 · 2012: 4
- FWCI
- 2.43
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Gastroenterology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Ministry of Education, Culture, Sports, Science and Technology
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- hepatic progenitor cells, fetal liver, serum-free culture, side population cells, Sca-1, CD49f, hepatocyte differentiation, cholangiocyte differentiation, liver regeneration, liver transplantation, mouse model
- MeSH terms
- Animals, Cell Survival, Flow Cytometry, Liver, Liver Regeneration, Mice, Inbred C57BL, Stem Cells, Time Factors, Antigens, CD, Culture Media, Serum-Free, Spheroids, Cellular, Stem Cell Transplantation, Cell Proliferation, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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