Enhanced Hepatogenic Differentiation of Human Wharton’s Jelly–Derived Mesenchymal Stem Cells by Using Three-Step Protocol

Abstract

Currently, human Wharton’s jelly-derived mesenchymal stem cells (hWJ-MSCs) are an attractive source of stem cells for cell-based therapy, owing to their ability to undergo self-renewal and differentiate into all mesodermal, some neuroectodermal, and endodermal progenies, including hepatocytes. Herein, this study aimed to investigate the effects of sodium butyrate (NaBu), an epigenetic regulator that directly inhibits histone deacetylase, on hepatic endodermal lineage differentiation of hWJ-MSCs. NaBu, at 1 mM, optimally promoted endodermal differentiation of hWJ-MSCs, along with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) supplementation (EGF + bFGF + 1 mM NaBu). CXCR4, HNF3β, SOX17 (endodermal), and GATA6 (mesendodermal) mRNAs were also up-regulated (p < 0.001). Immunocytochemistry and a Western blot analysis of SOX17 and HNF3β confirmed that the EGF + bFGF + 1 mM NaBu condition was appropriately pre-treated with hWJ-MSCs before hepatogenic differentiation. Furthermore, the hepatogenic medium + NaBu pre-treatment up-regulated hepatoblast (AFP and HNF3β) and hepatic (CK18 and ALB) markers, and increased the proportion of mature hepatocyte functions, including G6P, C/EBPα, and CYP2B6 mRNAs, glycogen storage and urea secretion. The hepatogenic medium + NaBu in the pre-treatment step can induce hWJ-MSC differentiation toward endodermal, hepatoblastic, and hepatic lineages. Therefore, the hepatogenic medium + NaBu pre-treatment for differentiating hWJ-MSCs could represent an alternative protocol for cell-based therapy and drug screening in clinical applications.

Small Molecule Treatments Improve Differentiation Potential of Human Amniotic Fluid Stem Cells

“…When cells were treated with small molecules individually, stable or upregulated cell viability was observed. Many papers report tendencies that agree with our results with decitabine ( Pang et al, 2019 ), TSA ( Han et al, 2013 ), NaBut ( Panta et al, 2019 ), RA ( Pourjafar et al, 2017 ), vitC ( Markmee et al, 2019 ). Decitabine is a well-known hypomethylating agent which incorporates itself into host DNA, but is used as a drug to treat myelodisplastic syndrome and acute myeloid leukemia.…”

Challenges for Deriving Hepatocyte-Like Cells from Umbilical Cord Mesenchymal Stem Cells forIn VitroToxicology Applications

“…However, the translation of these studies to UC-MSC is still limited. To the best of our knowledge, only Panta et al [26] showed that pre-treatment of UCM-MSCs with NaBu upregulated hepatoblast and hepatocyte markers and stimulated mature hepatic-associated functions, such as urea production, glycogen storage and G6P, CEBPA, and CYP2B6 activity, compared to non-treated differentiated cells.…”

“…Stepwise addition of factors such as EGF, FGF, HGF, nicotinamide, ITS, dexamethasone or OSM to the culture medium is used in the majority of the studies to differentiate MSCs [23,26,34,35,41]. Although the addition of these factors seems to lead to hepatic differentiation, a full mature hepatic cell has not yet been achieved.…”

Induction and Maturation of Hepatocyte-Like Cells In Vitro: Focus on Technological Advances and Challenges

“…The use of CHIR99021 reduce concentrations of activin A without affecting the differentiation rate of endoderm (Farzaneh et al, 2018). Sodium butyrate and valproic acid are histone deacetylation inhibitors that can promote the differentiation of definitive entoderm into liver-specific cells (Kondo et al, 2014;Panta et al, 2019). Trichostatin A, 5-aza, and nanomycin A, all of which are epigenetic modifiers, can be employed to induce differentiation of HLCs (Seeliger et al, 2013;Cipriano et al, 2017;Nakamae et al, 2018).…”