Journal: iScience

Article Title: AMPK-p38 axis converts human pluripotent stem cells to naive state

doi: 10.1016/j.isci.2026.115569

Figure Lengend Snippet: Naive conversion with AICAR (A) Naive conversion protocol using AICAR. PXGL; PD0325901 (1 μM), XAV939 (2 μM), Go6983 (2 μM), and human LIF (10 ng/mL) in Ndiff227 medium. (B) Flow cytometry analysis of EOS-GFP, SUSD2, CD75, and CD57 expression (day 14). (C) Appearance of EOS-GFP-positive cell clusters induced by AICAR (day 14). (D) EOS-GFP-positive naive-like colony after expansion in PXGL (day 14 + 14p). (E) Flow cytometry analysis of EOS-GFP, SUSD2, and CD75 expression (day 14 + 21p). (F) RT-qPCR analysis of sorted SUSD2 + CD75 + cells and parental primed human ESCs (H9-EOS). AICAR, day 14 + 13p; VPA, day 9 + 4p. Error bars: SD of technical triplicates. (G) Immunostaining for OCT4, NANOG, KLF17, and TFE3 of primed and AICAR-induced cells. AICAR, day 14 + 14p. (H) AICAR-treated cells showed increased TMRE fluorescence, as detected by both flow cytometry (PE channel) and fluorescence microscopy. Scale bars: 100 μm in (C, D, and H) and 50 μm (G). AICAR, day 14 + 16p. See also and and .

Article Snippet: Primed H9 human ESCs were obtained from WiCell (WA09).

Techniques: Flow Cytometry, Expressing, Quantitative RT-PCR, Immunostaining, Fluorescence, Microscopy

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: In vitro testing of R-28 cell-derived EVs on RGC survival and regeneration. Representative images of untreated control wells (A), R-28 cell-derived EVs treated wells (B), and CNTF-treated wells (C) are shown with the graphs showing the total surviving RGC number (D), the number of RGC with neurites (E), and the longest neurite length (F) in primary retinal cell culture after 3 days. Data are expressed as the mean ± SEM. Images were stained with a nuclear (DAPI, blue) and RGC marker (β-III tubulin, green). Scale bars: 50 µm. All experiments were performed in three independent biological replicates. CNTF: Ciliary neurotrophic factor; DAPI: 4′,6-diamidino-2-phenylindole; EV: extracellular vesicles; RGC: retinal ganglion cells.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: In Vitro, Derivative Assay, Control, Cell Culture, Staining, Marker

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: R-28 cell-derived EVs promote human ESC-derived RGC survival in vitro . Images show untreated controls, R-28 cell-derived EV treated, and CNTF-treated hESC-derived RGCs (green, βIII-tubulin) after injury induced by the microtubule poison, colchicine. Scale bar: 50 µm. Data are presented as mean ± SEM. All experiments were performed in three independent biological replicates. CNTF: Ciliary neurotrophic factor; ESC: embryonic stem cells; EV: extracellular vesicles; RGC: retinal ganglion cells.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: Derivative Assay, In Vitro

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: R-28 cell-derived EVs show protective trend for RGCs in a chronic glaucoma model. (A) Experimental design of the in vivo study. R-28 cell-derived EVs were intravitreally injected weekly beginning 1 week after microbead injection, and animals’ IOPs were measured twice a week. Four weeks after weekly EV injection, animals were sacrificed and histologically analyzed. After injection, microbeads localized (arrow) around the iridocorneal angle (B). IOP (mmHg) of healthy animals (blue) and animals receiving intracameral injection of microbeads with (green) or without (brown) intravitreal EV treatments is shown (C). (D, E) Representative images (D) and quantification (E) of Brn3a + (green) RGCs from the three groups on week 5. Scale bars: 50 µm. Data are presented as mean ± SEM. n = 3–5. EV: Extracellular vesicles; IOP: intraocular pressure; RGC: retinal ganglion cells; PBS: phosphate buffered saline.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: Derivative Assay, In Vivo, Injection, Saline

Journal: Neural Regeneration Research

Article Title: R-28 cell-derived extracellular vesicles protect retinal ganglion cells in glaucoma

doi: 10.4103/NRR.NRR-D-24-00709

Figure Lengend Snippet: Differentially expressed miRNA shown as abundance and fold change heat map profiles. Heatmaps show the upregulated and downregulated normalized counts of miRNA from injured RGCs treated with R-28 cell-derived EVs compared to injured untreated (A, B), injured RGCs treated with R-28 derived EVs compared to uninjured treated (D, E), and uninjured RGCs treated with R-28 cell-derived EVs compared to injured untreated (G, H), both statistically significant ( P < 0.05; A, D, G) and those trending towards significance ( P < 0.1; B, E, H) with abundance profiles shown in associated bar charts (C, F, I, respectively). * P < 0.05, ** P < 0.01. Data are presented as mean ± SEM. n = 3. EV: Extracellular vesicles; RGC: retinal ganglion cells.

Article Snippet: To test the therapeutic effect of R-28-derived EVs on human ESC-derived RGC (H7/H9 immortalized cell line; WiCell, Madison, WI, USA, #WA07, RRID: CVCL_S800) were differentiated from CRISPR-modified ESC generously donated from Prof Donald Zacks laboratory (Johns Hopkins University, Baltimore, MD, USA) and licensed for use from WiCell (Material Transfer Agreement issue-164634007).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Deep Learning-Guided Holotomography Reveals Early Structural Remodelling During Pluripotency Exit

doi: 10.64898/2026.04.23.720508

Figure Lengend Snippet: (a) Transcriptomic analysis of the time-course differentiation process. Loss of pluripotency markers is visible over time. (b) A GM25256 hiPSC colony after 12 h of RA-induced differentiation. The red arrow indicates filopodium-like membrane protrusion at colony periphery. The green arrow indicates an intercellular gap. (c) A GM25256 hiPSC colony after 24 h of RA-induced differentiation. (d) A GM25256 hiPSC colony after 48 h of RA-induced differentiation. The red arrow indicates a jagged colony boundary. ( e ) A GM25256 hiPSC colony after 96 h of RA-induced differentiation. The red arrow indicates an intercellular gap near the colony periphery. ( f ) A H9 hESC colony both untreated and after 24 h of RA-induced differentiation. P undiff was measured by DeepHOPE. (g) Ratio of H9 hESC colonies with average P undiff over 0.5. (h) A KOLF2.1J hiPSC colony both untreated and after 24 h of RA-induced differentiation. P undiff was measured by DeepHOPE. (i) Ratio of KOLF2.1J hiPSC colonies with average P undiff over 0.5. Scale bar (B to H) = 50 μm.

Article Snippet: The human embryonic stem cell line H9 (WA09, WiCell) and human iPSCs lines GM25256 (Coriell Institute) and KOLF2.1J (The Jackson Laboratory) were used to generate the base model for this study.

Techniques: Membrane