Table 1.
Characterization and validation.
| Classification | Test | Result | Data |
|---|---|---|---|
|
| |||
| Morphology | Photography bright field | Normal | Fig. 1A |
| Phenotype | Qualitative analysis | Positive expression of pluripotency markers by immunocytochemistry: NANOG, SOX2, and OCT3/4 | Fig. 1B |
| Genotype | Karyotype (G-banding) and resolution | Karyostat™ Assay, resolution 1–2 Mb: Normal karyotype 46, XY for both lines. | Fig. 1H |
| Identity | Microsatellite PCR (mPCR) or STR analysis | Not performed | N/A |
| 22 loci tested, 100% identical | Submitted in archive with journal | ||
| Mutation analysis (IF APPLICABLE) | Sequencing | SCVIi049-A: heterozygous c.25C > T | Fig. 1F |
| SCVIi050-A: heterozygous c.40_42delAGA | |||
| Southern blot OR WGS | Not performed | Not performed | |
| Microbiology and virology | Mycoplasma | Mycoplasma testing by luminescence: Negative (p10 and above) | Supplemental Fig. 1A |
| Differentiation potential | Embryoid body formation or Teratoma formation or Scorecard or directed differentiation | Directed differentiation, positive expression of germ layer markers | Fig. 1G |
| List of recommended germ layer markers | Expression of these markers has to be demonstrated at mRNA (RT PCR) or protein (IF) levels, at least 2 markers need to be shown per germ layer | Positive expression of germ layer markers: | Fig. 1G |
| Ectoderm: Pax6, Otx2 | |||
| Mesoderm: Brachyury, Tbx6 | |||
| Endoderm: Sox17, Foxa2 | |||
| Donor screening (OPTIONAL) | HIV 1 + 2 Hepatitis B, Hepatitis C | Not performed | Not performed |
| Genotype additional info (OPTIONAL) | Blood group genotyping | Not performed | Not performed |
| HLA tissue typing | Not performed | Not performed | |