surface stem cell marker tra1 60  (Miltenyi Biotec)

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: Stem Cells Translational Medicine

Article Title: Autophagy modulates tenogenic differentiation of cartilage-derived stem cells in response to mechanical tension via FGF signaling

doi: 10.1093/stcltm/szae085

Figure Lengend Snippet: Autophagy influences the tenogenic differentiation of cyclic tensile stress-treated CDSCs. (A) Flow cytometry analysis was performed to evaluate the expression of mouse mesenchymal stem cell-related markers CD90.2, CD44, CD29, and CD34 in P3 passage mouse CDSCs. (B) Time-lapse images show the morphological changes of CDSCs under cyclic tensile stress (5%, 1HZ). (C-D) PCR (C) and Western Blot (D) analyses were conducted to assess the expression levels of tenogenic and chondrogenic-related markers in CDSCs treated with cyclic tensile stress at different time points ( n = 3). T represents cyclic tension stress. (E) Western blot analysis was performed to examine autophagy-related marker expression in CDSCs treated with cyclic tensile stress at different time points. (F) Time-lapse images demonstrate the cellular morphology changes of CDSCs under both cyclic tensile stress and chloroquine treatment (40 μmol/L). (G-H) PCR (G) and Western Blot (H) analyses were carried out to determine the expression levels of tenogenic and chondrogenic-related markers in CDSCs subjected to both cyclic tensile stress and chloroquine treatment at different time points( n = 3). CQ denotes chloroquine. (I) Western blot analysis was conducted to investigate autophagy-related marker expression in CDSCs treated with cyclic tensile stress and chloroquine at different time points. Data are mean ± SD. *, P < .05; **, P < .01; ***, P < .001; ****, P < .0001.

Article Snippet: The characteristics of CDSCs were validated using the Mesenchymal Stem Cell (Mouse) Surface Marker Identification Kit (MUXMX-09011, Cyagen).

Techniques: Flow Cytometry, Expressing, Western Blot, Marker