Efficient maintenance of the undifferentiated status of human pluripotent stem cells (hiPSCs) is crucial for producing cells with improved proliferation, survival and differentiation, which can be successfully used for stem cell research and therapy. Here, we generated iPSCs from healthy donor peripheral blood mononuclear cells (PBMCs) and analyzed the proliferation and differentiation capacities of the generated iPSCs using single cell NGS-based 24-chromosome aneuploidy screening and RNA sequencing. In addition, we screened various natural compounds for molecules that could enhance the proliferation and differentiation potential of hiPSCs. Among the tested compounds, 3,2'-dihydroxyflavone (3,2'-DHF) significantly increased cell proliferation and expression of naïve stemness markers and decreased the dissociation-induced apoptosis of hiPSCs. Of note, 3,2'-DHF-treated hiPSCs showed upregulation of intracellular glutathione (GSH) and an increase in the percentage of GSH-high cells in an analysis with a FreSHtracer system. Interestingly, culture of the 3,2'-DHF-treated hiPSCs in differentiation media enhanced their mesodermal differentiation and differentiation into CD34+ CD45+ hematopoietic progenitor cells (HPC) and natural killer cells (NK) cells. Taken together, our results demonstrate that the natural compound 3,2'-DHF can improve the proliferation and differentiation capacities of hiPSCs and increase the efficiency of HPC and NK cell production from hiPSCs.

中文翻译：

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3,2'-Dihydroxyflavone改善人多能干细胞的增殖和存活及其向造血祖细胞的分化。

有效维持人类多能干细胞（hiPSC）未分化状态对于生产具有改善的增殖，存活和分化能力的细胞至关重要，而这些细胞可以成功用于干细胞的研究和治疗。在这里，我们从健康的供体外周血单核细胞（PBMC）生成了iPSC，并使用基于单细胞NGS的24染色体非整倍性筛选和RNA测序分析了生成的iPSC的增殖和分化能力。此外，我们从各种天然化合物中筛选了可以增强hiPSCs增殖和分化潜能的分子。在测试的化合物中，3,2'-二羟基黄酮（3,2' -DHF）显着增加了细胞增殖和幼稚干细胞标志物的表达，并降低了解离诱导的hiPSCs凋亡。值得注意的是，在使用FreSHtracer系统进行的分析中，经3,2'-DHF处理的hiPSC显示出细胞内谷胱甘肽（GSH）的上调和GSH高细胞百分比的增加。有趣的是，在分化培养基中培养3,2'-DHF处理的hiPSC增强了它们的中胚层分化，并分化为CD34 + CD45 +造血祖细胞（HPC）和自然杀伤细胞（NK）细胞。两者合计，我们的结果表明，天然化合物3,2'-DHF可以提高hiPSC的增殖和分化能力，并提高hiPSC产生HPC和NK细胞的效率。-DHF处理的hiPSC在FreSHtracer系统分析中显示出细胞内谷胱甘肽（GSH）的上调和GSH高细胞百分比的增加。有趣的是，在分化培养基中培养3,2'-DHF处理的hiPSC增强了它们的中胚层分化，并分化为CD34 + CD45 +造血祖细胞（HPC）和自然杀伤细胞（NK）细胞。两者合计，我们的结果表明，天然化合物3,2'-DHF可以提高hiPSC的增殖和分化能力，并提高hiPSC产生HPC和NK细胞的效率。-DHF处理的hiPSC在FreSHtracer系统分析中显示出细胞内谷胱甘肽（GSH）的上调和GSH高细胞百分比的增加。有趣的是，在分化培养基中培养3,2'-DHF处理的hiPSC增强了它们的中胚层分化，并分化为CD34 + CD45 +造血祖细胞（HPC）和自然杀伤细胞（NK）细胞。两者合计，我们的结果表明，天然化合物3,2'-DHF可以提高hiPSC的增殖和分化能力，并提高hiPSC产生HPC和NK细胞的效率。经DHF处理的hiPSC在分化培养基中增强了它们的中胚层分化，并分化为CD34 + CD45 +造血祖细胞（HPC）和自然杀伤细胞（NK）细胞。两者合计，我们的结果表明，天然化合物3,2'-DHF可以提高hiPSC的增殖和分化能力，并提高hiPSC产生HPC和NK细胞的效率。经DHF处理的hiPSC在分化培养基中增强了它们的中胚层分化，并分化为CD34 + CD45 +造血祖细胞（HPC）和自然杀伤细胞（NK）细胞。两者合计，我们的结果表明，天然化合物3,2'-DHF可以提高hiPSC的增殖和分化能力，并提高hiPSC产生HPC和NK细胞的效率。