Table 2.
Characterization and validation.
| Classification | Test | Result | Data |
|---|---|---|---|
| Morphology | Phase-contrast microscope | Normal | Fig. 1A |
| Phenotype | Qualitative analysis (immunofluorescence staining) | Expression of pluripotency markers: OCT4, NANOG, SSEA4 and TRA-1–60 | Fig. 1A |
| Quantitative analysis (RT-qPCR) | Expression of pluripotency markers: SOX2 and NANOG | Fig. 1E | |
| Qualitative analysis (FACS) | Expression of pluripotency markers: NANOG, SOX2, SSEA4, TRA-1–60, OCT4 | Fig. 1B | |
| Genotype | Karyotype (G-banding) and resolution | 46, XY; resolution 450–500 bands | Fig. 1F |
| Identity | Microsatellite PCR OR STR analysis | Not performed | N/A |
| 15 sites tested, 100% match | Supplementary file 1 | ||
| Mutation analysis (IF APPLICABLE) | DNA sequencing | Homozygous, OTULIN gene point mutation | Fig. 1D |
| Southern blot OR WGS | Not performed | N/A | |
| Microbiology and virology | Mycoplasma testing by luminescence | Negative | Supplementary file 2 |
| Differentiation potential | Monolayer differentiation assay | Differentiating cells are expression of RUNX1, AFP, and NESTIN; iPSC were able to differentiate into three germ layers | Fig. 1E |
| Donor screening (OPTIONAL) | HIV1 + HIV2, hepatitis B virus, hepatitis C virus | Not performed | N/A |
| Genotype additional info (OPTIONAL) | Blood group genotyping | Not performed | N/A |
| HLA tissue typing | Not performed | N/A |