Table 1.

Characterization and validation.

Classification	Test	Result	Data
Morphology	Photography	Typical human pluripotent stem cell morphology	Fig. 1B
Pluripotency status evidence for the described cell line	Qualitative analysis (Immunocytochemistry)	Positive for pluripotency markers: OCT3/4, NANOG, TRA1-60, SOX2	Fig. 1D
Karyotype	Karyotype (Wicell)	Normal male karyotype (46, XY), no clonal abnormalities detectedResolution: 425–450 bands	Fig. 1I
Genotyping for the desired genomic alteration/allelic status of the gene of interest	PCR across the edited site or targeted allele-specific PCR	PCR across integration site in edited and wild-type alleles.	Fig. 1C
	Transgene-specific PCR	N/A	N/A
Verification of the absence of random plasmid integration events	PCR/Southern	N/A	N/A
Parental and modified cell line genetic identity evidence	STR analysis (Wicell)	DNA Profiling 15 loci analyzed; matched to parental line.	Supplementary file submitted in the archive with journal
Mutagenesis/genetic modification outcome analysis	Sequencing (genomic DNA PCR or RT-PCR product)	Verified presence of integration in single edited allele and absence of integration in wild-type allele. Monoallelic transgene insertion.	Fig. 1F
	PCR-based analyses	Detection of correctly-targeted construct	Fig. 1C (Sample Lane 1)
	Southern Blot or WGS; western blotting (for knock-outs, KOs)	N/A	N/A
Off-target nuclease analysis-	PCR across top predicted likely off-target sites	PCR across predicted off-target site; Sanger sequencing	No off-target effect observed
Specific pathogen-free status	Mycoplasma testing by MycoAlert Detection Kit; passage 35	Negative	N/A
Multilineage differentiation potential	Spontaneous Differentiation; RNA isolation RNeasy kit (Qiagen); Taqman Scorecard (ThermoFisher Scientific)	Tri-lineage differentiation potential	Fig. 1G
Donor screening (OPTIONAL)	HIV 1 + 2 Hepatitis B, Hepatitis C	N/A	N/A
Genotype - additional histocompatibility info (OPTIONAL)	Blood group genotyping	N/A	N/A
	HLA tissue typing	N/A	N/A