Table 2.
Characterization and validation.
| Classification | Test | Result | Data |
|---|---|---|---|
| Morphology | Photography | Normal | Fig. 1 panel A |
| Phenotype | Qualitative analysis by immunocytochemistry | Immunocytochemistry for pluripotency markers OCT4, Nanog, SSEA4 and Tra-1–60 | Fig. 1 panel B |
| Quantitative analysis by RT-qPCR | qRT-PCR for expression of Nanog OCT4, REX1, SOX, HTERT and DNMT3B | Fig. 1 panel B | |
| Genotype | Karyotype (G-banding) and resolution | BCHNEUi001-A: 46,XY | Fig. 1 panel E |
| Band Resolution: 425–500 | |||
| BCHNEUi002-A: 46,XY | |||
| Band Resolution: 400–425 | |||
| BCHNEUi004-A: 46,XX | |||
| Band Resolution: 375–475 | |||
| BCHNEUi005-A: 46,XX | |||
| Band Resolution: 425–475 | |||
| BCHNEUi005-A: 46,XX | |||
| Band Resolution: 425–500 | |||
| BCHNEUi006-A 46,XX | |||
| Band Resolution: 450–500 | |||
| Identity | STR analysis | Performed | Archived with the journal |
| 16 loci tested, all matched (D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, PentaD, vWA, D8S1179, TPOX, FGA, Amelogenin) | Archived with the journal | ||
| Mutation analysis | Sanger sequencing | Confirmed variants listed in Table 1. | Fig. 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 (SNA1L2, RGS4 and HAND2) and endodermal markers (SST, KLF5 and AFP) | Fig. 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 |