Table 1.
Characterization and validation.
| Classification | Test | Result | Data |
|---|---|---|---|
| Morphology | Photography Bright field | Normal | Fig. 1A |
| Phenotype | Qualitative analysis (Immunocytochemistry) | Positive expression of pluripotency markers: Oct3/4, NANOG, SOX2 | Fig. 1B |
| Quantitative analysis (RT-qPCR) | mRNA expression of SOX2 and NANOG | Fig. 1D | |
| Genotype | Karyotype: Whole genome array (KaryoStat™ Assay) Resolution 1–2 Mb | Normal karyotype: 46 XX for BFVSBi003-A XX for BFVSBi004-A |
Fig. 1H |
| Identity | Microsatellite PCR (mPCR) or STR analysis | N/A | N/A |
| 16 loci tested, 100 % matching indentity | Submitted in archive with journal | ||
| Mutation analysis | Sequencing | COL3A1 gene (c.226A > G, p.Asn76Asp) | Fig. 1G |
| Southern Blot OR WGS | N/A | N/A | |
| Microbiology and virology | Mycoplasma | Luminescence: Negative | Fig. 1F |
| Differentiation potential | Directed differentiation, Immunofluorescence staining for 2 markers per germ layer | Positive Immunofluorescence staining of three germ layer markers Ectoderm: PAX6, OTX2 Endoderm: SOX17, FOXA2, Mesoderm: BRACHYURY, TBX6 |
Fig. 1C |
| Donor screening | HIV 1 + 2 Hepatitis B, Hepatitis C | N/A | N/A |
| Genotype additional info | Blood group genotyping | N/A | N/A |
| HLA tissue typing | N/A | N/A |