Table 2:
Characterization and Validation
| Classification | Test | Result | Data |
|---|---|---|---|
| Morphology | Photography | Normal | Figure 1 panel A |
| Phenotype | Qualitative analysis by immunocytochemistry | Immunocytochemistry for pluripotency markers OCT4, Nanog, SSEA4 and Tra-1-60 | Figure 1 panel B |
| Quantitative analysis by RT-qPCR | qRT-PCR for expression of Nanog, OCT4, REX1, SOX, HTERT and DNMT3B | Figure 1 panel B | |
| Genotype | Karyotype (G-banding) and resolution | BCHNEUi007-A: 46,XY Band Resolution: 425-475 BCHNEUi008-A: 46,XY Band Resolution: 375-475 BCHNEUi009-A: 46,XY Band Resolution: 450-525 BCHNEUi010-A: 46,XY Band Resolution: 475-525 BCHNEUi011-A: 46,XY Band Resolution: 425-475 BCHNEUi012-A: 46,XY Band Resolution: 500-550 |
Figure 1 panel E |
| Identity | STR analysis | Performed 16 loci tested, all matched (D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, Amelogenin) | Archived with the journal Archived with the journal |
| Mutation analysis | Sanger Sequencing | Confirmed variants listed in Table 1. | Figure 1 panel F |
| Southern Blot OR WGS | N/A | N/A | |
| Microbiology and virology | Mycoplasma | Mycoplasma testing by luminescence (MycoAlert™): Negative | Archived with the journal |
| Differentiation potential | Embryoid body formation | Expression of ectodermal (EN1, MAP2 and NR2F2), mesodermal (SNAIL2, RGS4 and HAND2) and endodermal markers (SST, KLF5 and AFP) | Figure 1 panel D |
| Donor screening (OPTIONAL) | HIV 1 + 2 Hepatitis B, Hepatitis C | N/A | N/A |
| Genotype additional info (OPTIONAL) | Blood group genotyping | N/A | N/A |
| HLA tissue typing | N/A | N/A |