Table 1. Human iPSC Line SCTi007-A Is Derived from a Male Donor with Early-Onset Parkinson’s Disease

Early-Onset Parkinson’s Disease Human iPSC Line, SCTi007-A, was derived from PBMCs of a 50-year-old male donor clinically diagnosed with Parkinson’s disease. Demographic, health, and genetic characteristics of the SCTi007-A donor are compiled based on self-reported information, clinical documentation and whole-exome sequencing. Sex was determined by karyotype. Ancestry and HLA haplotype were calculated from combined whole-genome and whole-exome sequencing data. Blood type (ABO/Rh blood group) was determined by next-generation sequencing. Height, weight, and BMI were calculated at the donation facility. iPSC = induced pluripotent stem cell; PBMC = peripheral blood mononuclear cell

Figure 1. Cells from Human iPSC Line SCTi007-A Exhibit High-Quality Morphology in Routine Culture

Cryopreserved cells from the Early-Onset Parkinson’s Disease Human iPSC Line, SCTi007-A, were thawed and maintained in mTeSR™ Plus on Corning® Matrigel® Matrix. Cells were cultured at 37°C and subsequently analyzed on Day 7 by brightfield microscopy. (A) The resulting iPSC colonies have densely packed cells and show multi-layering when ready to be passaged. (B,C) Cells retain prominent nucleoli and high nuclear-to-cytoplasmic ratios. iPSC = induced pluripotent stem cell

Figure 2. Human iPSC Line SCTi007-A Maintains a Normal Karyotype

(A) G-T-L banding for thawed cells from SCTi007-A at p20 (n = 20) shows a normal karyotype with no evidence of clonal abnormalities at a band resolution of 425 - 500 G-bands per haploid genome. (B) Fluorescent in situ hybridization in a representative p20 iPSC using probes for 20p11.21 (green) and 20q11.21 (red). 93% of cells examined displayed two sets of two probe signals, indicating no aneusomy of chromosome 20 (n = 200). iPSC = induced pluripotent stem cell

Table 2. Single Nucleotide Polymorphism Microarray Analysis Characterizes SCTi007-A Copy Number Variants

DNA was extracted from a vial of SCTi007-A iPSCs Commercial Cell Bank and subject to SNP microarray analysis to identify large-scale copy number variants (CNVs). The cells display two reportable genomic alterations, defined as copy number changes greater than 400kb or regions of loss of heterozygosity (LOH) greater than 5Mb. These include a 5.64Mb region of LOH on chromosome 1 and a 0.484Mb gain on chromosome 15 (rows highlighted in bold font). Array design, genomics position, genes, and chromosome banding are based on genome build GRCh37/hg19. chr = chromosome; bp = base pairs; SNP = single nucleotide polymorphism

Figure 3. Cells from Human iPSC Line SCTi007-A Express Undifferentiated Cell Markers

Cell line SCTi007-A was characterized using flow cytometry for undifferentiated cell markers OCT3/4 and TRA-1-60. (A) Percentage marker expression was quantified 3 passages after thawing from the analyses of two biological replicates. Representative flow cytometry plots are displayed for (B) OCT3/4 and (C) TRA-1-60. iPSC = induced pluripotent stem cell

Figure 4. Cells from Human iPSC Line SCTi007-A Demonstrate a High Trilineage Differentiation Capacity

Cells from SCTi007-A were split into 3 groups, differentiated using STEMdiff™ Trilineage Differentiation Kit (Catalog #05230), and then subjected to flow cytometry analysis. Two markers for each embryonic germ layer were assessed, and bars present mean marker expression for each group of cells (n = 2 biological replicates). PAX6 and Nestin confirm differentiation to the ectoderm lineage, NCAM and Brachyury (T) to the mesoderm lineage, and CXCR4 and SOX17 to the endoderm lineage

Table 3. Comprehensive Risk Variant Profile for the SCTi007-A iPSC Line Indicates a LRRK2 G2019S Parkinson’s Disease-Associated Mutation

The table presents risk variants identified for the SCTi007-A iPSC line. DNA was purified from the working cell bank, and whole-genome sequencing was performed to capture the entire genomic landscape. Sequencing was conducted on the NovaSeq X Plus System, achieving a 50X coverage. Coverage of coding regions and adjacent splice junction sites was enhanced for 20,000 genes using the SureSelect Human All Exon V6 Kit (Agilent Technologies). Variants were called and analyzed following GATK4 best practices, incorporating a Convolutional Neural Network to score each variant. High-confidence variants were annotated with predicted functional consequences and ClinVar IDs. ClinVar entries required two or more supporting ALT reads and an assertion criteria of one gold star or more in ClinVar to be considered. Heterozygous LRRK2 G2019S mutation was further confirmed via Sanger Sequencing. This comprehensive profile assists in understanding the genetic landscape and potential health impacts associated with the SCTi007-A iPSC line