Recent evidence suggests that progenitor cells in adult tissues and embryonic stem cells share a high resistance to hypoxia and ischemic stress. To study the ischemic resistance of adult liver progenitors, we characterized remaining viable cells in human liver tissue after cold ischemic treatment for 24–168 h, applied to the tissue before cell isolation. In vitro cultures of isolated cells showed a rapid decline of the number of different cell types with increasing ischemia length. After all ischemic periods, liver progenitor–like cells could be observed. The comparably small cells exhibited a low cytoplasm-to-nucleus ratio, formed densely packed colonies, and showed a hepatobiliary marker profile. The cells expressed epithelial cell adhesion molecule, epithelial-specific (CK8/18) and biliary-specific (CK7/19) cytokeratins, albumin, α-1-antitrypsin, cytochrome-P450 enzymes, as well as weak levels of hepatocyte nuclear factor-4 and γ-glutamyl transferase, but not α-fetoprotein or Thy-1. In vitro survival and expansion was facilitated by coculture with mouse embryonic fibroblasts. Hepatic progenitor–like cells exhibit a high resistance to ischemic stress and can be isolated from human liver tissue after up to 7 days of ischemia. Ischemic liver tissue from various sources, thought to be unsuitable for cell isolation, may be considered as a prospective source of hepatic progenitor cells.
Get full access to this article
View all access options for this article.
References
1.
RoskamsT.A., LibbrechtL., DesmetV.J.Progenitor cells in diseased human liver. Semin Liver Dis, 23:385. 2003.
2.
VesseyC.J., de la HallP.M.Hepatic stem cells: a review. Pathology, 33:130. 2001.
3.
TheiseN.D., SaxenaR., PortmannB.C., ThungS.N., YeeH., ChiribogaL., KumarA., CrawfordJ.M.The canals of Hering and hepatic stem cells in humans. Hepatology, 30:1425. 1999.
SchmelzerE.PottenC.S., ClarkeR.B., WilsonJ., RenehanA.G.Hepatic stem cellsthe liver's maturational lineages. Adult (Tissue) Stem Cells: Biology and Applications. New York: Taylor and Francis, 2006; 161–214.
6.
JangY.-Y., SharkisS.J.A low level of reactive oxygen species selects for primitive hematopoietic stem cells that may reside in the low-oxygenic niche. Blood, 110:3056. 2007.
7.
D'IppolitoG., DiabiraS., HowardG.A., MeneiP., RoosB.A., SchillerP.C.Marrow-isolated adult multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human cells with extensive expansion and differentiation potential. J Cell Sci, 117:2971. 2004.
8.
GraysonW.L., ZhaoF., BunnellB., MaT.Hypoxia enhances proliferation and tissue formation of human mesenchymal stem cells. Biochem Biophys Res Commun, 358:948. 2007.
9.
DanetG.H., PanY., LuongoJ.L., BonnetD.A., SimonM.C.Expansion of human SCID-repopulating cells under hypoxic conditions. J Clin Invest, 112:126. 2003.
10.
EzashiT., DasP., RobertsR.M.Low O2 tensions and the prevention of differentiation of hES cells. Proc Natl Acad Sci USA, 102:4783. 2005.
11.
SchmelzerE., WauthierE., ReidL.M.The phenotypes of pluripotent human hepatic progenitors. Stem Cells, 24:1852. 2006.
12.
SeldenC., ChalmersS.-A., JonesC., StandishR., QuagliaA., RolandoN., BurroughsA.K., RollesK., DhillonA., HodgsonH.J.F.Epithelial colonies cultured from human explanted liver in subacute hepatic failure exhibit hepatocyte, biliary epithelial, and stem cell phenotypic markers. Stem Cells, 21:624. 2003.
13.
WeiskirchenR., GressnerA.M.Isolation and culture of hepatic stellate cells. Methods Mol Med, 117:99. 2005.
14.
AlpiniG., LenziR., ZhaiW.R., LiuM.H., SlottP.A., ParonettoF., TavoloniN.Isolation of a nonparenchymal liver cell fraction enriched in cells with biliary epithelial phenotypes. Gastroenterology, 97:1248. 1989.
15.
OhS.K., KimH.S., ParkY.B., SeolH.W., KimY.Y., ChoM.S., KuS.Y., ChoiY.M., KimD.-W., MoonS.Y.Methods for expansion of human embryonic stem cells. Stem Cells, 23:605. 2005.
16.
LaursonJ., SeldenC., HodgsonH.J.F.Hepatocyte progenitors in man and in rodents—multiple pathways, multiple candidates. Int J Exp Pathol, 86:1. 2005.
17.
SellS.Electron microscopic identification of putative liver stem cells and intermediate hepatocytes following periportal necrosis induced in rats by allyl alcohol. Stem Cells, 15:378. 1997.
18.
WangJ., ClarkJ.B., RheeG.-S., FairJ.H., ReidL.M., GerberD.A.Proliferation and hepatic differentiation of adult-derived progenitor cells. Cells Tissues Organs, 173:193. 2003.
19.
McClellandR., WauthierE., UronisJ., ReidL.Gradients in the liver's extracellular matrix chemistry from periportal to pericentral zones: influence on human hepatic progenitors. Tissue Eng, 14:59. 2008.
20.
DanY.Y., RiehleK.J., LazaroC., TeohN., HaqueJ., CampbellJ.S., FaustoN.Isolation of multipotent progenitor cells from human fetal liver capable of differentiating into liver and mesenchymal lineages. Proc Natl Acad Sci USA, 103:9912. 2006.
21.
TurnerW.S., SchmelzerE., McClellandR., WauthierE., ChenW., ReidL.M.Human hepatoblast phenotype maintained by hyaluronan hydrogels. J Biomed Mater Res, 82:156. 2007.
22.
PetersenB.E., GoffJ.P., GreenbergerJ.S., MichalopoulosG.K.Hepatic oval cells express the hematopoietic stem cell marker Thy-1 in the rat. Hepatology, 27:433. 1998.
23.
WeissM.C., Strick-MarchandH.Isolation and characterization of mouse hepatic stem cells in vitro. Semin Liver Dis, 23:313. 2003.
24.
HerreraM.B., BrunoS., ButtiglieriS., TettaC., GattiS., DeregibusM.C., BussolatiB., CamussiG.Isolation and characterization of a stem cell population from adult human liver. Stem Cells, 24:2840. 2006.
25.
DuretC., Gerbal-ChaloinS., RamosJ., FabreJ.-M., JacquetE., NavarroF., BlancP., Sa-CunhaA., MaurelP., Daujat-ChavanieuM.Isolation, characterization, and differentiation to hepatocyte-like cells of nonparenchymal epithelial cells from adult human liver. Stem Cells, 25:1779. 2007.
26.
WeissT.S., LichtenauerM., KirchnerS., StockP., AurichH., ChristB., BrockhoffG., Kunz-SchughartL.A., JauchK.-W., SchlittH.J., ThaslerW.E.Hepatic progenitor cells from adult human livers for cell transplantation. Gut, 57:1129. 2008.
27.
ElmaouhoubA., DudasJ., RamadoriG.Kinetics of albumin- and alpha-fetoprotein-production during rat liver development. Histochem Cell Biol, 128:431. 2007.
28.
HarunaY., SaitoK., SpauldingS., NalesnikM.A., GerberM.A.Identification of bipotential progenitor cells in human liver development. Hepatology, 23:476. 1996.
29.
SicklickJ.K., ChoiS.S., BustamanteM., McCallS.J., PerezE.H., HuangJ., LiY.-X., RojkindM., DiehlA.M.Evidence for epithelial-mesenchymal transitions in adult liver cells. Am J Physiol Gastrointest Liver Physiol, 291:575. 2006.
30.
InadaM., FollenziA., ChengK., SuranaM., JosephB., BentenD., BandiS., QianH., GuptaS.Phenotype reversion in fetal human liver epithelial cells identifies the role of an intermediate meso-endodermal stage before hepatic maturation. J Cell Sci., 121:1002. 2008.
31.
AuthM.K., JoplinR.E., OkamotoM., IshidaY., McMasterP., NeubergerJ.M., BlahetaR.A., VoitT., StrainA.J.Morphogenesis of primary human biliary epithelial cells: induction in high-density culture or by coculture with autologous human hepatocytes. Hepatology, 33:519. 2001.
32.
CrosbyH.A., KellyD.A., StrainA.J.Human hepatic stem-like cells isolated using c-kit or CD34 can differentiate into biliary epithelium. Gastroenterology, 120:534. 2001.
33.
SimonM.C., KeithB.The role of oxygen availability in embryonic development and stem cell function. Nat Rev Mol Cell Biol, 9:285. 2008.
34.
WullschlegerS., LoewithR., HallM.N.TOR signaling in growth and metabolism. Cell, 124:471. 2006.
35.
KoumenisC., WoutersB.G.“Translating” tumor hypoxia: unfolded protein response (UPR)-dependent and UPR-independent pathways. Mol Cancer Res, 4:423. 2006.
36.
GrayJ.M., KarowD.S., LuH., ChangA.J., ChangJ.S., EllisR.E., MarlettaM.A., BargmannC.I.Oxygen sensation and social feeding mediated by a C. elegans guanylate cyclase homologue. Nature, 430:317. 2004.
37.
OhS.H., HatchH.M., PetersenB.E.Hepatic oval “stem” cell in liver regeneration. Semin Cell Dev Biol, 13:405. 2002.
38.
MitakaT., MizuguchiT., SatoF., MochizukiC., MochizukiY.Growth and maturation of small hepatocytes. J Gastroenterol Hepatol, 13 Suppl:S70. 1998.
39.
YasuiO., MiuraN., TeradaK., KawaradaY., KoyamaK., SugiyamaT.Isolation of oval cells from Long-Evans Cinnamon rats and their transformation into hepatocytes in vivo in the rat liver. Hepatology, 25:329. 1997.
40.
FujikawaT., HiroseT., FujiiH., OeS., YasuchikaK., AzumaH., YamaokaY.Purification of adult hepatic progenitor cells using green fluorescent protein (GFP)-transgenic mice and fluorescence-activated cell sorting. J Hepatol, 39:162. 2003.
41.
AzumaH., HiroseT., FujiiH., OeS., YasuchikaK., FujikawaT., YamaokaY.Enrichment of hepatic progenitor cells from adult mouse liver. Hepatology, 37:1385. 2003.
42.
SeldenC., HodgsonH.Cellular therapies for liver replacement. Transpl Immunol, 12:273. 2004.
43.
FiegelH.C., KaufmannP.M., BrunsH., KluthD., HorchR.E., VacantiJ.P., KneserU.Hepatic tissue engineering: from transplantation to customized cell-based liver directed therapies from the laboratory. J Cell Mol Med, 12:56. 2008.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
