Journal: Frontiers in Immunology
Article Title: Generation and characterization of human induced pluripotent stem cells from neuropathologically confirmed multiple system atrophy patient-derived fibroblasts
doi: 10.3389/fimmu.2026.1641981
Figure Lengend Snippet: Assessment of the cell line identity and the genomic integrity of the six MSA patient-derived iPSCs. (A) Short tandem repeat profiling provided the number of repeating units of 16 polymorphic microsatellite loci, to compare the donor fibroblast cells (reference) and the generated iPSCs and verify their origin. (B) G-banding revealed that the six iPSC lines showed a normal diploid karyotype (n = 46) and matched the sex of the donor patient. (C) Digital PCR allowed us to screen whether specific genomic regions accumulated duplications that may give a strong selective survival or growth advantage to iPSCs. Example of the scatter digital PCR plots displaying simultaneously (1) FAM signals, shown in purple and green to detect the target gene ( ID1 , NCAPD2 , PITX1 , RPS6KB1 , SOAT1 , or STS ), which are located in the 6 most recurrent abnormal regions; and (2) VIM signals, shown in orange and green to detect the reference gene, RPP30 . The ratio between the number of copies of the target gene and of the reference gene was used to determine the normal 2-copy number or copy number variations. The plots were obtained through QuantStudio 3D Digital PCR system. In all cases, we obtained 2-copy number (D) Quantitative Real Time RT-PCR was used for confirming the absence of the Sendai virus in the iPSC lines after 8 passages, through four primer sets (SeV, KOS, Klf4 and c-Myc). Box and Whisker plots depict in log10 scale the gene expression relative to GAPDH gene and normalized to the 7 days post-transduction fibroblasts, calculated by the 2(-ΔΔC(T)) method. In all iPSC clones, the expression levels of the SeV RNA were very low, reaching the limit of detection. Data are shown as the mean ± s.d. n = 3.
Article Snippet: SOAT1 , Hs06606603_cn , FAM , 1q25 , hg38|179355957-179356068 , 111 bp , HaeIII, HindIII, CviQI.
Techniques: Derivative Assay, Generated, Digital PCR, Quantitative RT-PCR, Virus, Whisker Assay, Gene Expression, Transduction, Clone Assay, Expressing