In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells

doi:10.1016/j.bbrc.2005.03.086

Biochemical and Biophysical Research Communications

Volume 330, Issue 4, 20 May 2005, Pages 1153-1161

https://doi.org/10.1016/j.bbrc.2005.03.086 Get rights and content

Abstract

In addition to long-term self-renewal capability, human mesenchymal stem cells (MSCs) possess versatile differentiation potential ranging from mesenchyme-related multipotency to neuroectodermal and endodermal competency. Of particular concern is hepatogenic potential that can be used for liver-directed stem cell therapy and transplantation. In this study, we have investigated whether human umbilical cord blood (UCB)-derived MSCs are also able to differentiate into hepatocyte-like cells. MSCs isolated from UCB were cultured under the pro-hepatogenic condition similar to that for bone marrow (BM)-derived MSCs. Expression of a variety of hepatic lineage markers was analyzed by flow cytometry, RT-PCR, Western blot, and immunofluorescence. The functionality of differentiated cells was assessed by their ability to incorporate DiI-acetylated low-density lipoprotein (DiI-Ac-LDL). As the cells were morphologically transformed into hepatocyte-like cells, they expressed Thy-1, c-Kit, and Flt-3 at the cell surface, as well as albumin, α-fetoprotein, and cytokeratin-18 and 19 in the interior. Moreover, about a half of the cells were found to acquire the capability to transport DiI-Ac-LDL. Based on these observations, and taking into account immense advantages of UCB over other stem cell sources, we conclude that UCB-derived MSCs retain hepatogenic potential suitable for cell therapy and transplantation against intractable liver diseases.

Section snippets

Materials and methods

Hepatic differentiation of UCB-derived MSCs. Processing of human full-term UCB samples and subsequent isolation of MSCs from UCB were accomplished as previously described [36], [37], [38]. Hepatic differentiation of UCB-derived MSCs was performed as described [4] with some modifications. Prior to hepatic differentiation, fifth passage cells were plated at a density of 1 × 10⁶ cells/cm² in culture medium and grown to 60% confluency. For initiation, the cells were incubated for 2 weeks in initiation

UCB-derived MSCs underwent morphological changes under the pro-hepatogenic condition

During the initiation step, the cells showed the remarkable transition from bipolar fibroblast-like morphology to round or oval shape (Fig. 1A). Right after exposure to initiation medium containing HGF, the cells began to lose their sharp edges and were progressively shrunk away, resulting in complete loss of the fibroblastic bipolar morphology. This morphological change was detected as early as the third day in the region of high cell density, and propagated into the area of low cell density,

Discussion

Stem cell-based therapy and transplantation are of potential value in tissue and organ replacement and regeneration approaches. Among various types of human stem cells, MSCs have been regarded as one of the most promising “off-the-shelf” cell therapeutics for the following reasons. First, isolation and ex vivo expansion of MSCs are relatively facile so that they can be obtained in quantity appropriate for clinical application. Second, a large number of different donor cells bearing human

Acknowledgments

This research was supported in part by a grant (SC3200) from Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Science and Technology and a grant (10012112) from Components and Materials Technology Development Program funded by the Ministry of Commerce, Industry and Energy, Republic of Korea.

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In vitro differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocyte-like cells

Abstract

Section snippets

Materials and methods

UCB-derived MSCs underwent morphological changes under the pro-hepatogenic condition

Discussion

Acknowledgments

FEBS Lett.

Hepatology

Hepatology

Differentiation

Hepatology

Am. J. Pathol.

Biochem. Biophys. Res. Commun.

Blood

Biochem. Biophys. Res. Commun.

Blood

Gastroenterology

Blood

Biol. Blood Marrow Transplant.

Biochem. Biophys. Res. Commun.

Cytotherapy

Biochem. Biophys. Res. Commun.

Hepatology

Blood

Cytokine

Hepatology

Mesenchymal stem cells: use and perspectives

Hematol. J.

Mesenchymal stem cells

Exp. Biol. Med.

Neural cells derived from adult bone marrow and umbilical cord blood

J. Neurosci. Res.

In vitro hepatic differentiation of human mesenchymal stem cells

Hepatology

Differentiation of pancreatic epithelial progenitor cells into hepatocytes following transplantation into rat liver

Proc. Natl. Acad. Sci. USA