Abstract
Although stem cells are commonly isolated by fluorescence-activated cell sorting or magnetic affinity cell sorting, they are very expensive, and they need known markers. However, there is no specific marker for liver stem/progenitor cells (LSPCs). Here, we describe a convenient and efficient method (three-step method) to enrich LSPCs. The fetal liver cells (FLCs) were firstly enriched by Percoll discontinuous gradient centrifugation from the rat fetal liver. Then the FLCs in culture were purified to be homogeneous in size by differential trypsinization and differential adherence. Finally, fetal liver stem/progenitor cells (FLSPCs) were enriched from purified FLCs by Percoll continuous gradient centrifugation. Flow cytometric analysis combining with marker CD133 was used to detect the purity of FLSPCs and evaluate the isolating effects of the three-step method.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Schmelzer E et al (2007) Human hepatic stem cells from fetal and postnatal donors. J Exp Med 204:1973–1987
Oertel M et al (2006) Cell competition leads to a high level of normal liver reconstitution by transplanted fetal liver stem/progenitor cells. Gastroenterology 130(2):507–520; quiz 90
Greenbaum LE, Wells RG (2011) The role of stem cells in liver repair and fibrosis. Int J Biochem Cell Biol 43(2):222–229
Russo FP, Parola M (2011) Stem and progenitor cells in liver regeneration and repair. Cytotherapy 13(2):135–144
Oertel M et al (2008) Purification of fetal liver stem/progenitor cells containing all the repopulation potential for normal adult rat liver. Gastroenterology 134(3):823–832
Nierhoff D et al (2007) New cell surface markers for murine fetal hepatic stem cells identified through high density complementary DNA microarrays. Hepatology 46(2):535–547
Liu WH, Li R, Dou KF (2011) Convenient and efficient enrichment of the CD133+ liver cells from rat fetal liver cells as a source of liver stem/progenitor cells. Stem Cell Rev 7(1):94–102
Yu S et al (2004) Isolation and characterization of the CD133+ precursors from the ventricular zone of human fetal brain by magnetic affinity cell sorting. Biotechnol Lett 26(14):1131–1136
Hao HN et al (2003) Fetal human hematopoietic stem cells can differentiate sequentially into neural stem cells and then astrocytes in vitro. J Hematother Stem Cell Res 12(1):23–32
Shmelkov SV et al (2005) Cytokine preconditioning promotes codifferentiation of human fetal liver CD133+ stem cells into angiomyogenic tissue. Circulation 111(9):1175–1183
Kordes C et al (2007) CD133+ hepatic stellate cells are progenitor cells. Biochem Biophys Res Commun 352(2):410–417
Overturf K et al (1999) The repopulation potential of hepatocyte populations differing in size and prior mitotic expansion. Am J Pathol 155(6):2135–2143
Sandhu JS et al (2001) Stem cell properties and repopulation of the rat liver by fetal liver epithelial progenitor cells. Am J Pathol 159(4):1323–1334
Rajvanshi P et al (1996) Studies of liver repopulation using the dipeptidyl peptidase IV-deficient rat and other rodent recipients: cell size and structure relationships regulate capacity for increased transplanted hepatocyte mass in the liver lobule. Hepatology 23(3):482–496
Nagai H et al (2002) Differentiation of liver epithelial (stem-like) cells into hepatocytes induced by coculture with hepatic stellate cells. Biochem Biophys Res Commun 293(5):1420–1425
Alpini G et al (1994) Recent advances in the isolation of liver cells. Hepatology 20(2):494–514
Piscaglia AC et al (2010) Stem cell-based therapies for liver diseases: state of the art and new perspectives. Stem Cells Int 2010:259461
Khan AA et al (2010) Human fetal liver-derived stem cell transplantation as supportive modality in the management of end-stage decompensated liver cirrhosis. Cell Transplant 19(4):409–418
Acknowledgments
Our study was supported by the National Natural Science Foundation of China (NO30772102). We thank Fu-qin Zhang (the Fourth Military Medical University, China), who gave us a lot of help in the experimental design. Sincere thanks also go to Rui Guo (Sichuan International Studies of University, China) for language organizing.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Liu, W., You, N., Dou, K. (2012). Convenient and Efficient Enrichment of the CD133+ Liver Cells from Rat Fetal Liver as a Source of Liver Stem/Progenitor Cells. In: Singh, S. (eds) Somatic Stem Cells. Methods in Molecular Biology, vol 879. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-815-3_9
Download citation
DOI: https://doi.org/10.1007/978-1-61779-815-3_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-814-6
Online ISBN: 978-1-61779-815-3
eBook Packages: Springer Protocols