Embryo Technology and Stem Cell Research Center, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
School of Pathology and Laboratory Medicine, Institute of Medicine, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand.
Int J Mol Sci. 2019 Jun 20;20(12):3016. doi: 10.3390/ijms20123016.
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). , , (endodermal), and (mesendodermal) mRNAs were also up-regulated ( < 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 , , and 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.
目前，人沃顿氏胶源间充质干细胞（hWJ-MSCs）因其具有自我更新能力并能分化为所有中胚层、部分神经外胚层和内胚层祖细胞，包括肝细胞，因此是细胞治疗中一种有吸引力的干细胞来源。本研究旨在探讨组蛋白去乙酰化酶直接抑制剂丁酸钠（NaBu）对 hWJ-MSCs 肝内胚层谱系分化的影响。1mM 的 NaBu 与表皮生长因子（EGF）和碱性成纤维细胞生长因子（bFGF）补充剂（EGF+bFGF+1mM NaBu）一起最佳地促进 hWJ-MSCs 的内胚层分化。实时定量 RT-PCR 分析显示，与对照组相比，内胚层标志物（SOX17、GATA4、FOXA2、HNF4α）和中胚层标志物（T 基因、MIXL1）mRNA 的表达水平上调（<0.001）。免疫细胞化学和 Western blot 分析 SOX17 和 HNF3β 证实，EGF+bFGF+1mM NaBu 条件下 hWJ-MSCs 可在肝向分化前进行适当的预处理。此外，肝向培养基+NaBu 预处理上调肝前体细胞（AFP 和 HNF3β）和肝（CK18 和 ALB）标志物，并增加成熟肝细胞功能的比例，包括细胞色素 P450 相关基因（CYP3A4、CYP2B6、UGT1A1）、葡萄糖储存和尿素分泌。肝向培养基+NaBu 预处理可诱导 hWJ-MSC 向内胚层、肝前体细胞和肝谱系分化。因此，肝向培养基+NaBu 预处理分化 hWJ-MSCs 可能代表细胞治疗和临床应用中药物筛选的替代方案。
