Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells
Robert E. Schwartz, … , Wei-Shou Hu, Catherine M. Verfaillie
Robert E. Schwartz, … , Wei-Shou Hu, Catherine M. Verfaillie
Published May 15, 2002
Citation Information: J Clin Invest. 2002;109(10):1291-1302. https://doi.org/10.1172/JCI15182.
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Multipotent adult progenitor cells from bone marrow differentiate into functional hepatocyte-like cells

Abstract

We have derived from normal human, mouse, and rat postnatal bone marrow primitive, multipotent adult progenitor cells (MAPCs) that can differentiate into most mesodermal cells and neuroectodermal cells in vitro and into all embryonic lineages in vivo. Here, we show that MAPCs can also differentiate into hepatocyte-like cells in vitro. Human, mouse, and rat MAPCs, cultured on Matrigel with FGF-4 and HGF, differentiated into epithelioid cells that expressed hepatocyte nuclear factor-3β (HNF-3β), GATA4, cytokeratin 19 (CK19), transthyretin, and α-fetoprotein by day 7, and expressed CK18, HNF-4, and HNF-1α on days 14–28. Virtually all human, as well as a majority of rodent cells stained positive for albumin and CK18 on day 21; 5% (rodent) to 25% (human) cells were binucleated by day 21. These cells also acquired functional characteristics of hepatocytes: they secreted urea and albumin, had phenobarbital-inducible cytochrome p450, could take up LDL, and stored glycogen. MAPCs, which can be expanded in vitro and maintained in an undifferentiated state for more than 100 population doublings, can thus differentiate into cells with morphological, phenotypic, and functional characteristics of hepatocytes. MAPCs may therefore be an ideal cell for in vivo therapies for liver disorders or for use in bioartificial liver devices.

Authors

Robert E. Schwartz, Morayma Reyes, Lisa Koodie, Yuehua Jiang, Mark Blackstad, Troy Lund, Todd Lenvik, Sandra Johnson, Wei-Shou Hu, Catherine M. Verfaillie

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Quantitative RT-PCR and Western blot analyses confirm hepatocyte-like ph...
Quantitative RT-PCR and Western blot analyses confirm hepatocyte-like phenotype. (a) mMAPCs and (b) hMAPCs were cultured on Matrigel with FGF-4 and HGF or FGF-4 alone for 21 and 28 days, respectively. At multiple time points during culture, cells were harvested and underwent quantitative RT-PCR using the SYBR green method for mRNAs as indicated. The mRNA levels were normalized using β-actin as a housekeeping gene. Numbers under the blots for CK18, CK19, TTR, albumin, and CYP1B1 represent mRNA levels after treatment relative to undifferentiated MAPCs. For αFP, Cyp2b9, and Cyp2b13, numbers under the blots are relative to mRNA from the liver, because no transcripts were detected in undifferentiated MAPCs. Li, mouse or human liver mRNA; NT, no template. Representative example of five mouse and one human study. (c) hMAPCs (from b) were cultured on Matrigel with FGF-4 and HGF or FGF-4 alone for 21 days. HUH7 cells were cultured as described. Lysates of cells were separated by SDS-PAGE, transferred to Immuno-Blot PVDF membrane, and incubated for 1 minute with Ab’s against αFP, CK18, albumin, or β-actin. FN, FGF-4–induced hMAPCs on FN; M, FGF-4–induced hMAPCs on Matrigel; FH, FGF-4– and HGF-induced hMAPCs on Matrigel; Huh, Huh7 cell line used as control.