Journal: bioRxiv

Article Title: Replicable generation of rhesus macaque iPSCs for in vitro modeling of genetic frontotemporal dementia

doi: 10.64898/2026.03.17.712482

Figure Lengend Snippet: Effect of RhiPSC culture conditions on cloning efficiency. (A) Representative images of alkaline phosphatase-stained colonies from RhiPSC line M6 clone 16.02 on mouse embryonic fibroblasts (MEFs), Matrigel, or Vitronectin in UPPS medium with either no supplement, Y-27632, or CEPT. Scale bar, 100 µm. (B) Graph depicting percent (%) cloning efficiency (y-axis) of alkaline phosphatase-positive colonies cultured on different matrices (x-axis) in UPPS medium with no supplement (circles), Y-27632 (squares), or CEPT (triangles). Individual data points correspond to the mean (dashed lines) % cloning efficiency of RhiPSC colonies counted among six technical replicates per cell line (M4 clone 14.09, red; M6 clone 16.02, pink; W4 clone 14.06, black; W6 clone 5.07, grey). Black brackets indicate statistically significant differences between two supplement types within a substrate group; black lines indicate statistically significant differences between two substrates with the same supplement. p < 0.05 is considered statistically significant. Tukey’s multiple comparisons post-hoc p-values: a=0.0488, b=0.0098, c=0.0097, d=0.0133, e=0.0177, f=0.0483, g=0.0166.

Article Snippet: After 5 days, colonies were stained with a Vector Blue Alkaline Phosphatase Substrate kit (Vector Laboratories, cat. no. SK-5300) as described in .

Techniques: Cloning, Staining, Cell Culture

Journal: bioRxiv

Article Title: Replicable generation of rhesus macaque iPSCs for in vitro modeling of genetic frontotemporal dementia

doi: 10.64898/2026.03.17.712482

Figure Lengend Snippet: Characterization of RhiPSC line W6 clone 5.07. (A) Amplicon sequencing of MAPT R406W with wild-type C allele indicated in the black box with a red arrow compared to reference genome (ref). (B) Phase contrast image of colonies grown on MEFs and (C) positive alkaline phosphatase staining; scale bar, 100 µm. (D) RT-PCR detection of endogenous pluripotent mRNAs POU5F1 , SOX2 , NANOG , KLF4 , LIN28 , and MYC . ACTB was used as a reference gene. (E) Amplification of EBNA and oriP in RhiPSCs at P16, no template control (NTC), and the rhesus embryonic stem cell line r420 (RhESC). Plasmid EM2K was used as a positive control (PC) for the detection of EBNA and oriP. (F) Positive immunofluorescence staining for pluripotency proteins SOX2 (green), OCT4 (red), and NANOG (far red), colocalized with DAPI (blue). Scale bar, 100 µm. (G) Flow cytometry-detected APC-A+ cells were 94.5% positive for SOX2, 99.2% positive for OCT4, and 95% positive for NANOG. (H) Short tandem repeat (STR) analysis confirmed (+) RhiPSC line identity. Mycoplasma (Myco.), Herpes B Virus (HBV), Simian Immunodeficiency Virus (SIV), Simian T-Lymphotropic Virus (STLV), and Simian Retrovirus (SRV) were not detected (-). (I) Normal 42,XX karyotype at P12. (J) H&E staining of a teratoma containing endoderm, mesoderm, and ectoderm. Scale bar, 100 µm.

Article Snippet: After 5 days, colonies were stained with a Vector Blue Alkaline Phosphatase Substrate kit (Vector Laboratories, cat. no. SK-5300) as described in .

Techniques: Amplification, Sequencing, Staining, Reverse Transcription Polymerase Chain Reaction, Control, Plasmid Preparation, Positive Control, Immunofluorescence, Flow Cytometry, Virus

Journal: bioRxiv

Article Title: Replicable generation of rhesus macaque iPSCs for in vitro modeling of genetic frontotemporal dementia

doi: 10.64898/2026.03.17.712482

Figure Lengend Snippet: Characterization of RhiPSC line M6 clone 16.02. (A) Amplicon sequencing of MAPT R406W with mutant T allele indicated in the black box with a red arrow compared to reference genome (ref). (B) Phase contrast image of colonies grown on MEFs and (C) positive alkaline phosphatase staining; scale bar, 100 µm. (D) RT-PCR detection of endogenous pluripotent mRNAs POU5F1 , SOX2 , NANOG , KLF4 , LIN28 , and MYC . ACTB was used as a reference gene. (E) Amplification of EBNA and oriP in RhiPSCs at P18, no template control (NTC), and the rhesus embryonic stem cell line r420 (RhESC). Plasmid EM2K was used as a positive control (PC) for the detection of EBNA and oriP. (F) Positive immunofluorescence staining for pluripotency proteins SOX2 (green), OCT4 (red), and NANOG (far red), colocalized with DAPI (blue). Scale bar, 100 µm. (G) Flow cytometry-detected APC-A+ cells were 93.1% positive for SOX2, 98% positive for OCT4, and 93.3% positive for NANOG. (H) Short tandem repeat (STR) analysis confirmed (+) RhiPSC line identity. Mycoplasma (Myco.), Herpes B Virus (HBV), Simian Immunodeficiency Virus (SIV), Simian T-Lymphotropic Virus (STLV), and Simian Retrovirus (SRV) were not detected (-). (I) Normal 42,XX karyotype at P15. (J) H&E staining of a teratoma containing endoderm, mesoderm, and ectoderm. Scale bar, 100 µm.

Article Snippet: After 5 days, colonies were stained with a Vector Blue Alkaline Phosphatase Substrate kit (Vector Laboratories, cat. no. SK-5300) as described in .

Techniques: Amplification, Sequencing, Mutagenesis, Staining, Reverse Transcription Polymerase Chain Reaction, Control, Plasmid Preparation, Positive Control, Immunofluorescence, Flow Cytometry, Virus

Journal: International Journal of Molecular Sciences

Article Title: Development of a High-Efficacy Reprogramming Method for Generating Human Induced Pluripotent Stem (iPS) Cells from Pathologic and Senescent Somatic Cells

doi: 10.3390/ijms21186764

Figure Lengend Snippet: TRA-1-60 and alkaline phosphatase staining of iPS cell colonies induced from the dermal tissue of three elderly patients. ( a ) TRA-1 immunofluorescent staining of iPS cell colonies. Immunofluorescent staining with anti-TRA1-60 antibody was performed on D1 (top), D2 (middle), and D3 (bottom) cells subjected to protocol 1 (left) or protocol 2 (right) for iPS cell induction at day 35 post-transfection. An integration of 81 continuous fields of the view captured with the Keyence BZ-X710 microscope is shown (observed area: 23.915 mm × 17.936 mm). BF, bright field. Arrows indicate TRA1-60-positive iPS cell colonies. Scale bars: 3.0 mm. ( b ) Alkaline phosphatase staining of iPS cell colonies. Staining with alkaline phosphatase was performed on D1 (top), D2 (middle), and D3 (bottom) cells subjected to protocol 1 (left) or protocol 2 (right) for iPS cell induction at day 36 post-transfection. Arrows indicate alkaline phosphatase-positive iPS cell colonies. ( c ) The number of iPS cell colonies stained by alkaline phosphatase was quantitively analyzed in all fibroblasts. The left side is protocol 1 and the right side is protocol 2. Values shown are the mean ± SEM (* p < 0.05, n = 3).

Article Snippet: An Anti-TRA-1-60 antibody, anti-SSEA4 antibody, and alkaline phosphatase stain kit (AP Staining Kit, Red-Color) was purchased from Funakoshi ® (Tokyo, Japan).

Techniques: Staining, Transfection, Microscopy