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Remodeling of Glycosaminoglycans During Differentiation of Adult Human Bone Mesenchymal Stromal Cells toward Hepatocytes.
Stem Cells and Development 2018 December 21
There is a critical need to generate functional hepatocytes to aid in liver repair and regeneration upon availability of a renewable, and potentially personalized, source of human hepatocytes. Currently, the vast majority of primary human hepatocytes are obtained from human tissue through cadavers. Recent advances in stem cell differentiation have opened up the possibility to obtain fully functional hepatocytes from embryonic or induced pluripotent stem cells, or adult stem cells. With respect to the latter, human bone marrow mesenchymal stromal cells (hBMSCs) can serve as a source of autogenetic and allogenic multipotent stem cells for liver repair and regeneration. A major aspect of hBMSC differentiation is the extracellular matrix (ECM) composition, and in particular, the role of glycosaminoglycans (GAGs) in the differentiation process. In the current study, we examine the influence of four distinct culture conditions/protocols (T1-T4) on GAG composition and hepatic markers. -Fetoprotein and hepatocyte nuclear factor (HNF)-4 were expressed continually over 21 days of differentiation, as indicated by RT-qPCR analysis, while albumin expression did not begin until day 21. Hepatocyte growth factor (HGF) appears to be more effective than activin A in promoting hepatic-like cells through the mesenchymal-epithelial transition, perhaps due to the former binding to the HGF receptor to form a unique complex that diversifies the biological functions of HGF. Of the four protocols tested, uniform hepatocyte-like morphological changes, albumin secretion, and glycogen storage were found to be highest with protocol T2, which involves both early and late stage combinations of growth factors. The total GAG profile of the hBMSC ECM is rich in heparan sulfate and hyaluronan, both of which fluctuate during differentiation. The GAG profile of primary human hepatocytes showed a heparan sulfate (HS)-rich ECM, and thus, it may be possible to guide hBMSC differentiation to more mature hepatocytes by controlling the GAG profile expressed by differentiating cells.
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