Journal: Stem Cell Research & Therapy

Article Title: Activation of the G-protein coupled estrogen receptor 1 (GPER1) reduces transient receptor potential vanilloid 1 (TRPV1) activity and human iPSC-derived nociceptive neuron firing

doi: 10.1186/s13287-026-05174-3

Figure Lengend Snippet: Reprogramming of adult human dermal fibroblasts (HDFa) into induced pluripotent stem cell (iPSC) line BO-VC1. Reprogramming was performed using the Epi5 ™ Episomal iPSC Reprogramming Kit, enabling the generation of ( A ) transgene- and virus-free iPSC line BO-VC1. Successful generation of fibroblast-derived iPSCs was validated by immunocytochemical staining for the pluripotency markers ( B ) SOX2, C TRA 1–60, D OCT4, SSEA4 and E NANOG. Quantification of the generated iPSCs via flow cytometry revealed F 99.31% SSEA4/TRA1-60, G 97.43% SOX2/TRA1-60 and H 98.60% OCT3/4/TRA1-60 positive cells. I Quantification of the transcript levels of the stem cell markers NANOG , OCT4 , REX1 and SOX2 revealed significantly higher mRNA levels in iPSC line BO-VC1 compared to the HDFa control ( n = 3 different passages) Moreover, generated iPSCs were functionally validated by directed differentiation into all three germ layers. Subsequent immunocytochemical staining of the iPSCs before and after differentiation confirmed the expression of ( J–M ) meso-, N–Q endo- and R–U ectodermal lineage markers only in the respective differentiations. Scale bars: 50 μm. Data were tested for normal distribution using Shapiro-Wilk test. Means ± SEM (standard error of the mean) were statistically analyzed by a Kruskal-Wallis test with Dunn’s multiple comparisons test. n = 3 (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001)

Article Snippet: After harvesting, using Accutase, 1 × 10 6 cells were stained for the surface stem cell marker TRA1-60 (1:50, #130-122-965, Miltenyi Biotec, Bergisch Gladbach, Germany) and SSEA4 (1:50, #130-124-073, Miltenyi Biotec, Bergisch Gladbach, Germany) in 25 μl PEB buffer (PBS + 0.5% BSA) for 10 min at 4 °C, washed in 500 μl PEB buffer and centrifuged at 200xg for 5 min.

Techniques: Virus, Derivative Assay, Staining, Generated, Flow Cytometry, Control, Expressing

Journal: bioRxiv

Article Title: Microfluidic Mechanical Reactivation of Aged Stem Cells

doi: 10.64898/2026.02.25.707893

Figure Lengend Snippet: (A) Representative fluorescence images and comparative analysis of intracellular ROS levels among P6, P11, P18, and P18 μ-CPR cells. Scale bar = 500 µm. (B) Representative fluorescence images showing intracellular ROS levels in P18 μ-CPR under three flow conditions ( Re = 89, 267, and 446). Scale bar = 500 µm. (C) RT-qPCR analysis of stemness-related genes ( OCT4 , SOX2 , and KLF4 ) after 3 days. Expression levels are normalized to P7 WJ-MSCs and presented as relative ratios for the P18 μ-CPR groups ( Re = 89, 267, and 446). (D) Representative immunofluorescence images showing the expression of stemness markers (OCT4, SOX2, and SSEA4) in P18 and P18 μ-CPR cells. Nuclei were stained with DAPI. Scale bar = 20 µm. (E–G) Quantification of fluorescence intensity (fold change) for OCT4 (E), SOX2 (F) , and SSEA4 (G) in P18 and P18 μ-CPR cells. (H) Quantification of Western blot band intensities (fold change) for stemness markers (OCT4, SOX2, and SSEA4) and the proliferation marker PCNA in P18 μ-CPR cells relative to the P18 control. (I) Immunoblotting of OCT4, SOX2, SSEA4, PCNA, and β-actin in μ-CPR and control MSCs. (J) Relative quantification of senescent cells using SA-β-Gal staining in P6, P11, P18, and P18 μ-CPR cells. The graph presents the percentages of SA-β-Gal-positive cells. Scale bar = 500 µm. (K, L) Evaluation of intracellular γH2AX expression and its quantification in P6, P11, P18, and P18 μ-CPR cells. Scale bar = 20 µm. Data are presented as the mean ± SEM of three independent experiments. Statistical analysis was performed using one-way ANOVA (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Specimens were incubated overnight at 4 °C in PBST with 1% bovine serum albumin and the following primary antibodies: OCT4 (sc-9081; Santa Cruz Biotechnology), SOX2 (sc-20088; Santa Cruz Biotechnology), SSEA4 (sc-21704; Santa Cruz Biotechnology), phospho-histone H2A.X (2577S; Cell Signaling Technology), actin (sc-8432; Santa Cruz Biotechnology), α-actinin (sc-17829; Santa Cruz Biotechnology), and α-tubulin (sc-23948; Santa Cruz Biotechnology).

Techniques: Fluorescence, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Western Blot, Marker, Control, Quantitative Proteomics