Journal: Molecular Therapy Advances

Article Title: Ferrostatin-1 and hinokitiol supplementation enhance human hematopoietic stem cell expansion in a chemically defined medium

doi: 10.1016/j.omta.2026.201711

Figure Lengend Snippet: Effect of antioxidant compounds on human CB CD34 + cell expansion and immunophenotype (A) Relative proliferation of CB CD34 + cells at day 14 in 3a medium supplemented with each of the compounds in SCREEN-WELL REDOX library. Data from three independent experiments, each performed with unique CB donor, are represented as relative luminescence (%) to DMSO-treated cells. Each row represents a compound. Well ID is shown on the left. (B and C) Fold change in HSPC (B) and HSC (C) fraction under each treatment condition relative to DMSO control group at day 14 of ex vivo culture. Representative data from 3 independent experiments, each performed with unique CB donor.

Article Snippet: Human CB CD34 + cells were purchased from HemaCare (CB34C-3) and STEMCELL Technologies (200-0001).

Techniques: Control, Ex Vivo

Journal: Molecular Therapy Advances

Article Title: Ferrostatin-1 and hinokitiol supplementation enhance human hematopoietic stem cell expansion in a chemically defined medium

doi: 10.1016/j.omta.2026.201711

Figure Lengend Snippet: Effect of selected compounds on CD34 + cell expansion in 3a medium Human CB CD34 + cells were seeded in 96-well plates at 10,000 cells per well in 3a medium with selected compounds. (A) Relative cell proliferation (fold increase) in the presence of selected compounds at day 14 compared to untreated control cells. Cell proliferation was assessed using CellTiter-Glo Luminescent cell viability assay. (B) Immunophenotypic analysis showing HSC percentage within HSPCs. (C) Lipid peroxidation levels at day 14 following treatment with selected compounds analyzed by BODIPY 581/591 C11 staining. Data show BODIPY-ox negative portion (%) in HSPCs. (D) Cellular ROS levels at day 14 following treatment with selected compounds measured with CellROX Deep Red. Data show negative portion (%) in HSPCs. Representative data from three independent experiments, each performed with unique CB donor in duplicate, are shown. (B–D) Mean ± SD, statistical analyses were conducted between Fer-1 versus DMSO vehicle control by t test, ∗ p < 0.05.

Article Snippet: Human CB CD34 + cells were purchased from HemaCare (CB34C-3) and STEMCELL Technologies (200-0001).

Techniques: Control, Cell Viability Assay, Staining

Journal: Molecular Therapy Advances

Article Title: Ferrostatin-1 and hinokitiol supplementation enhance human hematopoietic stem cell expansion in a chemically defined medium

doi: 10.1016/j.omta.2026.201711

Figure Lengend Snippet: Effect of selected compounds and Fer-1 combination on human CB CD34 + cell expansion Human CB CD34 + cells were seeded in 96-well plates at 10,000 cells per well in 3a medium with selected compounds with or without Fer-1. (A) Relative cell proliferation (fold change) at day 14 compared to untreated cells. Pooled data from three independent experiments, each performed with unique CB donor cells, are shown. (B and C) HSPC (B) and HSC (C) percentage in live cells with indicated compounds with or without Fer-1. Data represent three independent experiments, each performed with unique CB donor cells.

Article Snippet: Human CB CD34 + cells were purchased from HemaCare (CB34C-3) and STEMCELL Technologies (200-0001).

Techniques:

Journal: Molecular Therapy Advances

Article Title: Ferrostatin-1 and hinokitiol supplementation enhance human hematopoietic stem cell expansion in a chemically defined medium

doi: 10.1016/j.omta.2026.201711

Figure Lengend Snippet: Effect of Fer-1 and hinokitiol combination (FHK) on human CB CD34 + cell expansion Human CB CD34 + cells were seeded in 6-well plates at 300,000 cells per well in 3a medium with DMSO (vehicle control), or Fer-1 (10 μM) plus hinokitiol (0.5 μM) (FHK). (A) Relative cell proliferation at day 14 compared to DMSO. Pooled data from 3 independent experiments performed in duplicate. Mean ± SEM, ∗∗ p < 0.01 by t test. (B and C) Immunophenotypic analysis showing percentage of human HSPC (B) and HSC (C) in ex vivo expanded CB CD34 + cells at day 14. Pooled data from 3 independent experiments with CD34 + cells from 4 unique CB donors, performed in duplicate. (D and E) Lipid peroxidation (D) and intracellular ROS (E) levels in ex vivo expanded CD34 + CD45RA − cells measured by BODIPY 581/591 C11 and CellROX Deep Red staining, respectively. Representative overlaid histograms (left) and mean fluorescence intensity (MFI, right) are shown. Pooled data from 2 independent experiments with CD34 + cells from 2 unique CB donors, performed in duplicate. (F and G) Colony forming unit (CFU) activity in ex vivo expanded human CB CD34 + cells cultured for 14 days. Colony count of total progenitors (F) and various types of colonies (G) were quantified. Pooled data from 3 independent experiments with CD34 + cells from 4 unique CB donors, performed in duplicate. Mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗∗ p < 0.0001 by one-way ANOVA with Tukey’s multiple-comparison test except (G). Each CB donor is represented by a unique symbol.

Article Snippet: Human CB CD34 + cells were purchased from HemaCare (CB34C-3) and STEMCELL Technologies (200-0001).

Techniques: Control, Ex Vivo, Staining, Fluorescence, Activity Assay, Cell Culture, Comparison

Journal: Molecular Therapy Advances

Article Title: Ferrostatin-1 and hinokitiol supplementation enhance human hematopoietic stem cell expansion in a chemically defined medium

doi: 10.1016/j.omta.2026.201711

Figure Lengend Snippet: Expanded human CB cells engraftment and chimerism after transplantation (A) Schematic outlining the xenotransplantation experiment in NOG-EXL mice. Schematic created in BioRender.com. (B) Human blood cell chimerism (CD45 + cell percentage) in peripheral blood at indicated time point (left: pooled data, right: individual mouse data. (C–E) Human blood cell lineage distribution (chimerism ratio) at week 24 in peripheral blood (C), spleen (D), and bone marrow (E) of transplanted mice. Pooled data from two independent experiments performed with expanded CD34 + cells from two unique CB donors (represented by unique symbol ● and▲). Fresh cells from each donor were used as control for comparison. Experiment#1, N = 3 mice per group. Experiment#2, N = 4 (fresh), 4(DMSO), and 5(FHK). Mean ± SEM, two-way ANOVA with Tukey’s multiple-comparison test (right). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. (F) Bone marrow analyses at week 24 post-transplantation showing human CD45 + cells, lineage - cells, HSPCs, and HSC distribution. Data from one transplantation experiment are shown. Mean ± SEM, one-way ANOVA with Tukey’s multiple-comparison test. ∗∗∗ p < 0.001.

Article Snippet: Human CB CD34 + cells were purchased from HemaCare (CB34C-3) and STEMCELL Technologies (200-0001).

Techniques: Transplantation Assay, Control, Comparison