Table 1. iPSC Line SCTi005-A Is Derived from a Healthy Female Donor

Demographic, health, and genetic characteristics of the SCTi005-A donor are compiled based on self-reported information and whole-exome sequencing. Sex was determined by karyotype. Ancestry and HLA haplotype were calculated from whole-genome and whole-exome sequencing combined 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.

Figure 1. SCTi005-A Human Pluripotent Stem Cells Demonstrate High-Quality Morphology in Routine Culture

Cryopreserved cells from line SCTi005-A were thawed and maintained in mTeSR™ Plus on Corning® Matrigel® Matrix.

(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. SCTi005-A Human Pluripotent Stem Cells Maintain a Normal Karyotype

(A) G-T-L banding for thawed cells at p12 (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 p12 iPSC using probes for 20p11 (green) and 20q11.21 (red). 95% 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 SCTi005-A Copy Number Variants

DNA was extracted from a vial of SCTi005-A from the Master Cell Bank and subject to SNP microarray analysis to identify large-scale copy number variants (CNVs). The cells display no reportable CNVs, defined as those greater than 400kb in size. Array design, genomics position, genes, and chromosome banding are based on genome build GRCh38/hg3. chr = chromosome; bp = base pairs; SNP = single nucleotide polymorphism.

Figure 3. SCTi005-A iPSCs Express Undifferentiated Cell Markers

Cell line SCTi005-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. SCTi005-A Human Pluripotent Stem Cells Demonstrate a High Trilineage Differentiation Capacity

Cells from SCTi005-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 SCTi005-A iPSC Line

The table presents risk variants identified for the SCTi005-A iPSC line. DNA was purified from the master cell bank, and whole-genome sequencing was performed to capture the entire genomic landscape. Sequencing was conducted on the NovaSeq 6000 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. This comprehensive profile assists in understanding the genetic landscape and potential health impacts associated with the SCTi005-A iPSC line.