Differentiation of human adipose stromal cells into hepatic lineage in vitro and in vivo

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Abstract

Embryonic stem cells (ES cells), bone marrow-derived mesenchymal stem cells, umbilical cord blood-derived mesenchymal stem cells, and hepatic stem cells in liver have been known as a useful source that can induce to differentiate into hepatocytes. In this study, we examined whether human adipose tissue-derived stromal cells (hADSC) can differentiate into hepatic lineage in vitro. hADSC, that were induced to differentiate into hepatocyte-like cells by the treatment of HGF and OSM, had morphology similar to hepatocytes. Addition of DMSO enhanced differentiation into hepatocytes. RT-PCR and immunocytochemical analysis showed that hADSC express albumin and α-fetoprotein during differentiation. Differentiated hADSC showed LDL uptake and production of urea. Additionally, transplanted hADSC to CCl4-injured SCID mouse model were able to be differentiated into hepatocytes and they expressed albumin in vivo. Mesenchymal stem cells isolated from human adipose tissue are immunocompatible and are easily isolated. Therefore, hADSC may become an alternative source to hepatocyte regeneration or liver cell transplantation.

Section snippets

Isolation and culture of hADSC and HepG2 cell line

hADSC culture. After informed consent, leftover adipose tissues were obtained from four different donors (19 years old; male, 28 years old; female, 36 years old; female, and 55 years old; male) undergoing elective abdominoplasty. To isolate hADSC, adipose tissues were washed with equal volumes of phosphate-buffered saline (PBS), and tissues were digested at 37 °C for 30 min with 0.075% collagenase type I (Sigma). Enzyme activity was neutralized with α-modified Eagle’s medium (α-MEM), containing

Morphologic changes in cultured hADSC treated with HGF, OSM or DMSO

To specify antigenic properties of hADSC, we determined surface protein expression by flow cytometry. hADSC expressed CD29, CD44, CD105, and CD90, but did not express CD34, CD45, CD14, and HLA-DR, as described in our previous study [26]. To determine optimal conditions for hADSC differentiation into hepatocyte-like cells, we tested the effect of different cytokines on morphological changes of hADSC in the fibronectin-coated dishes. Differentiation was induced at 90% confluency of cells after

Discussion

MSC could differentiate into cells of all mesodermal origin, including adipocytes, osteocytes, chondrocytes, myocytes, and endothelial cells [20], [21], [22], [23]. Besides these, MSC are also capable of “transdifferentiation” into ectodermal cells, such as neural cells [27], [28]. These findings suggest that MSC belong to multipotent adult stem cells. Schwartz et al. [11] isolated a non-hematopoietic stem cell subset (CD45GlyA in humans or CD45Ter119 in mice) from bone marrow, termed

Acknowledgment

This work was supported by a grant (02-PJ10-PG8-EC01-0018) from the Ministry of Health and Welfare.

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