Table 2:
Characterization and validation
| Classification | Test | Result | Data |
|---|---|---|---|
| Morphology | Photography | All iPSC lines are morphologically normal | Not shown; available from authors. |
| Phenotype | Qualitative analysis | Confirmed expression of pluripotency markers: OCT4, NANOG, and SOX2 | Figure 1, panel B |
| Quantitative analysis | Assessed % of positive cells for pluripotency cell surface marker, SSEA-4. All lines are more than 98% positive. | Figure 1 panel B | |
| Genotype | Karyotype (G-banding) and resolution | 1. 46XX, Resolution 425–450 2. 46XX, Resolution 425–525 3. 46XX, Resolution 400–425 4. 46XX, Resolution 425–450 5. 46XX, Resolution 375–475 6. 46XX, Resolution 400–450 7. 46XX, Resolution 400–450 8. 46XX, Resolution 450–475 |
Figure 1 panel A |
| Identity | Microsatellite PCR (mPCR) OR STR analysis |
STR analysis | Supplementary data 1 |
| 16 specific loci were tested. All isogenic lines matched 100%. | Submitted in archive with journal | ||
| Mutation analysis (IF APPLICABLE) | Sequencing | 1. A53T 40760 heterozygous | Figure 1 panel C |
| 2. Iso-WT 40760 PL3B3 3. Iso-WT 40760 PL1E11 4. Iso-WT 40760 PL3B7 5. A53T 42072 heterozygous 5. Iso-WT 42072 PL2B9 6. Iso-WT 42072 PL2C11 7. Iso-WT 42072 PL3G1 |
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| Southern Blot OR WGS | Not performed | N/A | |
| Microbiology and virology | Mycoplasma | Mycoplasma testing was done by RT-PCR. All clones-negative | Supplementary figure 1 |
| Differentiation potential | e.g. Embryoid body formation OR Teratoma formation OR Scorecard OR Directed differentiation |
The STEMdiff™ Trilineage Differentiation Kit (StemCell) was used to test differentiation potential. We confirmed the expression of endoderm (SOX-17), mesoderm (Brachyury) and ectoderm ( -Tubulin III) markers in all clones. |
Figure 1 panel D |
| Donor screening (OPTIONAL) | HIV 1 + 2 Hepatitis B, Hepatitis C | Not performed | N/A |
| Genotype additional info (OPTIONAL) | Blood group genotyping | Not performed | N/A |
| HLA tissue typing | Not performed | N/A |