Journal: Stem cell research & therapy

Article Title: Footprint-free induced pluripotent stem cells can be successfully differentiated into mesenchymal stromal cells in the feline model.

doi: 10.1186/s13287-025-04325-2

Figure Lengend Snippet: Fig. 2 Feline iPSCs express pluripotent characteristics, pluripotency markers, and silencing of exogenous transgenes. (A). Characteristic morphology of established iPSC colonies after being passaged onto feeder cells. Arrows indicate cells with a high nuclear-to-cytoplasmic ratios. (B & C). Conventional RT-PCR analysis of endogenously expressed feline pluripotency markers SOX2, NANOG and OCT4, and the loading control GAPDH, in feline iPSCs at pas sage (P) 0, 3, 6, 9 and 12 (B) and P15 and 25 (C). Full-length gels are presented in Additional File 9: Fig. 9). (D). Conventional RT-PCR analysis of human exogenous transcription factors c-Myc, KOS, KLF4, and Sendai virus (SV), in feline iPSCs at P 0, 3, 6, 9 and 12. Feline GAPDH was included as loading control. (E). Expression of SeV using qPCR analysis at different passages

Article Snippet: Generation of feline iPSCs using retroviral (RV) vectors Retroviral plasmids based on the Moloney murine leukemia virus (MMLV) were purchased from Addgene and contained the coding sequences for the human transcription factors: OCT4 (Addgene #17217), SOX2 (Addgene #17218), NANOG (Addgene #18115), c-MYC (Addgene #17220), and KLF4 (Addgene #17219).

Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Virus, Expressing

Journal: Stem cell research & therapy

Article Title: Footprint-free induced pluripotent stem cells can be successfully differentiated into mesenchymal stromal cells in the feline model.

doi: 10.1186/s13287-025-04325-2

Figure Lengend Snippet: Fig. 3 Alkaline phosphatase staining, karyotype analysis, and embryoid body (EB) formation assay. (A). Feline iPSC colonies were stained for alkaline phos phatase or left unstained (control). (B). Karyotype analysis of feline iPSCs-1 and iPSCs − 2 at P20 showing a normal diploid chromosome number of 38. (C). Feline iPSCs formed embryoid bodies (EBs) in suspension culture in differentiation medium. (D). Conventional RT-PCR analysis of feline markers of all three embryonic germ layers, including alpha-fetoprotein (AFP), GATA binding protein 6 (GATA6), and C-X-C chemokine receptor type 4 (CXCR4) for endoderm; smooth muscle actin (SMA) and GATA2 for mesoderm; and ENOLASE and NESTIN for ectoderm in feline EBs. Feline GAPDH was included as loading control. Full-length gels are presented in Additional File 10: Fig. S10. (E). Conventional RT-PCR analysis of endogenously expressed feline pluripotency markers OCT4, SOX2, and NANOG, and the loading control GAPDH, in feline iPSCs and EBs

Article Snippet: Generation of feline iPSCs using retroviral (RV) vectors Retroviral plasmids based on the Moloney murine leukemia virus (MMLV) were purchased from Addgene and contained the coding sequences for the human transcription factors: OCT4 (Addgene #17217), SOX2 (Addgene #17218), NANOG (Addgene #18115), c-MYC (Addgene #17220), and KLF4 (Addgene #17219).

Techniques: Staining, Tube Formation Assay, Control, Suspension, Reverse Transcription Polymerase Chain Reaction, Binding Assay

Journal: Stem cell research

Article Title: Generation of human induced pluripotent stem cell line from a patient with a long QT syndrome type 2.

doi: 10.1016/j.scr.2015.12.039

Figure Lengend Snippet: Fig. 1. Characterization of human iPS cell line UKKi009-A. A. Morphology of iPS cell colonies as observed by bright field microscopy (left image: 10×, right image: 32×). B. Immunostaining for OCT4, NANOG, TRA-1-80 and SSEA4 showing the iPS colonies uniformly expressing the markers. C. Expression of pluripotent stem cell markers TRA-1-80 and SSEA4 on the surface of UKKi009-A iPS cells as measured by flow cytometry. D. Assessment of pluripotency of iPS cells by RT-qPCR analysis of expression of indicated germ layer-specific transcripts at day 14 of differentiation compared against undifferentiated cells as a control using the ΔΔCt method. E. RT-PCR showing absence of transgenes SOX2 and c-MYC (left panel) and OCT4 and KLF4 (right panel) expression in iPS cells. Plasmid encoding OSKM cassette was used as a positive control. Arrows indicate the expected band size (1848 bp and 1364 bp for left and right panel, respectively). M = DNA-size marker. F. DNA sequence showing the presence of mutation c.3035-3045delTCCCTCGATGC in the KCNH2 gene of patient-derived dermal fibroblasts (hDFs) and UKKi009-A iPS cells derived from them. The nucleotide sequence indicated beneath the sequencing chromatogram is the NCBI Reference Sequence: XM_011516185.1. G. Single nucleotide polymorphism (SNP) genotyping reveals identity of UKKi009-A iPS cells with NP0011 patient-derived hDFs abut not with hDFs derived from an unrelated donor NP0022.

Article Snippet: The cells were then stained overnight at 4 °C with primary antibodies specific for OCT4 (Santa Cruz Biotechnology, Cat. No. Sc-5279, 1:100), NANOG (R&D Systems, Cat. No. AF1997, 1:100) TRA1-80 (Santa Cruz, Cat. No. Sc-21706, 1:200) and SSEA4 (Santa Cruz, Cat. No. Sc-21704, 1:800).

Techniques: Microscopy, Immunostaining, Expressing, Cytometry, Quantitative RT-PCR, Control, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Positive Control, Marker, Sequencing, Mutagenesis, Derivative Assay