3,2′-Dihydroxyflavone Improves the Proliferation and Survival of Human Pluripotent Stem Cells and Their Differentiation into Hematopoietic Progenitor Cells

Abstract

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.

Improved Isolation and Culture of Urine-Derived Stem Cells (USCs) and Enhanced Production of Immune Cells from the USC-Derived Induced Pluripotent Stem Cells

“…Given that differentiation capacity into HPCs of USC-iPSC is lower than that of PBMC-iPSCs, 3,2 -DHF and 3,4 -DHF treatment could be utilized to overcome the limitation of USC-iPSC's differentiation capacity into HPCs. We also observed that flavonoid enhanced the population of CD34+/CD45+ cells in PBMC-iPSC [43]. Overall, these data suggest that flavonoids, 3,2 -DHF and 3,4 -DHF could possibly facilitate the efficient use of USCs by enhancing the efficiencies of USC isolation and differentiation of USC-iPSCs into HPCs and further into hematopoietic lineage cells.…”

“…Human dermal fibroblasts (HDFs) with passage 5, adipose-derived stem cells (ADSC) with passage 1, and Wharton's jelly-derived MSCs (WJ-MSCs) with passage 2 were obtained from Cell2in (Seoul, Republic of Korea) (IRB 7001355-201507-BR-181). The human peripheral blood mononuclear cells (PBMCs) were prepared from the blood of healthy donors using density gradient centrifugation via Ficoll-Paque PLUS (GE Healthcare, Chicago, IL, USA) (IRB 7001355-201507-BR-072) as we described [43]. Basically, the USCs isolation from human urine samples were carried out based on the previously established protocol [44].…”

“…Finally, the iPSC medium was replaced with mTeSR1 until colonies formed. We previously reported the detail of PBMC-iPSC clones [43].…”

Immunomodulatory effect of mesenchymal stem cells and mesenchymal stem-cell-derived exosomes for COVID-19 treatment

“…Recently, we devised an improved protocol for the isolation of urine-derived stem cells and their further differentiation into immune cells ( 95 ). Moreover, our research group promoted the hematopoietic differentiation of hiPSCs using a novel small molecule ( 96 ). At the advent of COVID-19, it has become mandatory to discover therapeutic strategies that are easily reproducible and cost effective.…”

pH dependency of the structural and photophysical properties of the atypical 2′,3-dihydroxyflavone

“…29 It protects cells against oxidative damage, 30,31 and is currently studied for its ability to improve the preparation and isolation of stem cells for regenerative medicine applications. [32][33][34] Its crystal structure has been characterized, using X-rays, 35 and its mass spectrum recorded. 36 2 0 3HF is a good candidate of study, as it is a simple 3HF derivative, but has shown to behave quite differently from other avonols.…”