List of patients or healthy subjects from whom iPSCs were generated
| Healthy donor or patient with: . | Age (y) . | CD34+ cell purification . | Starting CD34+ cell or blood amount . | TRA-1-60+ colonies . | Number Cre-excised . | Number assessed for 3-germ layer . |
|---|---|---|---|---|---|---|
| STEMCCA-loxP generated iPSC from apheresis collected G-CSF mobilized peripheral blood CD34+ cells | ||||||
| Healthy (iNC-01) | 44 | MACS | 20 000 cells | 4 | 1 | 1 |
| X-CGD (iGP91-07) | 21 | MACS | 50 000 cells | 10 | 1 | 1 |
| X-SCID (iSCID-01) | 17 | Dynal | 50 000 cells | 4 | 1 | 0 |
| STEMCCA-loxP generated iPSCs from nonmobilized peripheral blood CD34+cells | ||||||
| Healthy (iNC-02) | 42 | Dynal | 20 mL | 5 | 1 | 0 |
| Healthy (iNC-03) | 46 | Dynal | 20 mL | 5 | 1 | 0 |
| Healthy (iNC-04) | 52 | MACS | 14 mL | 7 | 1 | 0 |
| Healthy (iNC-05) | 38 | MACS | 28 mL | 3 | 1 | 0 |
| X-CGD (iGP91-01) | 42 | Dynal | 20 mL | 6 | 1 | 1 |
| X-CGD (iGP91-02) | 39 | Dynal | 20 mL | 2 | 1 | 1 |
| X-CGD (iGP91-03) | 36 | MACS | 20 mL | 2 | 2 | 2 |
| X-CGD (iGP91-04) | 30 | MACS | 18 mL | 1 | 1 | 1 |
| X-CGD (iGP91-05) | 12 | MACS | 20 mL | 5 | 0 | 0 |
| AR-CGD (iP22-01) | 19 | MACS | 20 mL | 10 | 1 | 0 |
| AR-CGD (iP47-01) | 37 | Dynal | 20 mL | 5 | 1 | 1 |
| AR-CGD (iP47-02) | 38 | Dynal | 20 mL | 9 | 1 | 1 |
| AR-CGD (iP47-03) | 30 | Dynal | 20 mL | 6 | 1 | 1 |
| AR-CGD (iP47-04) | 23 | MACS | 30 mL | 4 | 0 | 0 |
| AR-CGC (iP67-01) | 51 | MACS | 25 mL | 5 | 1 | 0 |
| X-SCID (iSCID-02) | 17 | MACS | 10 mL | 4 | 1 | 0 |
| X-SCID (iSCID-03) | 19 | MACS | 30 mL | 10 | 1 | 0 |
| X-CGD (iGP91-06) | 16 | MACS | Cryopreserved PBMC 20 mL | 3 | 1 | 0 |
| X-CGD (iGP91-08) | 52 | MACS | Cryopreserved PBMC 20 mL | 6 | 0 | 0 |
| AR-CGD (iP67-02) | 5 | MACS | Cryopreserved PBMC 10 mL | 1 | 1 | 0 |
| Sendai vectors generated iPSCs from nonmobilized peripheral blood CD34+ cells | ||||||
| Healthy (iNC-04-s) | 52 | MACS | 6 mL | >120 (8) | n/a | 2 |
| Healthy (iNC-06-s) | 51 | MACS | 5 mL | 3 (3) | n/a | 0 |
| X-CGD (iGP91-09-s) | 34 | MACS | 9 mL | 10 (5) | n/a | 0 |
| X-CGD (iGP91-10-s) | 22 | MACS | 4 mL | 274 (12) | n/a | 0 |
| X-CGD (iGP91-11-s) | 25 | MACS | 9 mL | 17 (8) | n/a | 0 |
| X-CGD (iGP91-12-s) | 2 | MACS | 2.5 mL | 408 (24) | n/a | 0 |
| Healthy donor or patient with: . | Age (y) . | CD34+ cell purification . | Starting CD34+ cell or blood amount . | TRA-1-60+ colonies . | Number Cre-excised . | Number assessed for 3-germ layer . |
|---|---|---|---|---|---|---|
| STEMCCA-loxP generated iPSC from apheresis collected G-CSF mobilized peripheral blood CD34+ cells | ||||||
| Healthy (iNC-01) | 44 | MACS | 20 000 cells | 4 | 1 | 1 |
| X-CGD (iGP91-07) | 21 | MACS | 50 000 cells | 10 | 1 | 1 |
| X-SCID (iSCID-01) | 17 | Dynal | 50 000 cells | 4 | 1 | 0 |
| STEMCCA-loxP generated iPSCs from nonmobilized peripheral blood CD34+cells | ||||||
| Healthy (iNC-02) | 42 | Dynal | 20 mL | 5 | 1 | 0 |
| Healthy (iNC-03) | 46 | Dynal | 20 mL | 5 | 1 | 0 |
| Healthy (iNC-04) | 52 | MACS | 14 mL | 7 | 1 | 0 |
| Healthy (iNC-05) | 38 | MACS | 28 mL | 3 | 1 | 0 |
| X-CGD (iGP91-01) | 42 | Dynal | 20 mL | 6 | 1 | 1 |
| X-CGD (iGP91-02) | 39 | Dynal | 20 mL | 2 | 1 | 1 |
| X-CGD (iGP91-03) | 36 | MACS | 20 mL | 2 | 2 | 2 |
| X-CGD (iGP91-04) | 30 | MACS | 18 mL | 1 | 1 | 1 |
| X-CGD (iGP91-05) | 12 | MACS | 20 mL | 5 | 0 | 0 |
| AR-CGD (iP22-01) | 19 | MACS | 20 mL | 10 | 1 | 0 |
| AR-CGD (iP47-01) | 37 | Dynal | 20 mL | 5 | 1 | 1 |
| AR-CGD (iP47-02) | 38 | Dynal | 20 mL | 9 | 1 | 1 |
| AR-CGD (iP47-03) | 30 | Dynal | 20 mL | 6 | 1 | 1 |
| AR-CGD (iP47-04) | 23 | MACS | 30 mL | 4 | 0 | 0 |
| AR-CGC (iP67-01) | 51 | MACS | 25 mL | 5 | 1 | 0 |
| X-SCID (iSCID-02) | 17 | MACS | 10 mL | 4 | 1 | 0 |
| X-SCID (iSCID-03) | 19 | MACS | 30 mL | 10 | 1 | 0 |
| X-CGD (iGP91-06) | 16 | MACS | Cryopreserved PBMC 20 mL | 3 | 1 | 0 |
| X-CGD (iGP91-08) | 52 | MACS | Cryopreserved PBMC 20 mL | 6 | 0 | 0 |
| AR-CGD (iP67-02) | 5 | MACS | Cryopreserved PBMC 10 mL | 1 | 1 | 0 |
| Sendai vectors generated iPSCs from nonmobilized peripheral blood CD34+ cells | ||||||
| Healthy (iNC-04-s) | 52 | MACS | 6 mL | >120 (8) | n/a | 2 |
| Healthy (iNC-06-s) | 51 | MACS | 5 mL | 3 (3) | n/a | 0 |
| X-CGD (iGP91-09-s) | 34 | MACS | 9 mL | 10 (5) | n/a | 0 |
| X-CGD (iGP91-10-s) | 22 | MACS | 4 mL | 274 (12) | n/a | 0 |
| X-CGD (iGP91-11-s) | 25 | MACS | 9 mL | 17 (8) | n/a | 0 |
| X-CGD (iGP91-12-s) | 2 | MACS | 2.5 mL | 408 (24) | n/a | 0 |
Shown in each column starting from the left column for each iPSC derivation are: (1) Whether the donor is a healthy donor or has a type of CGD or X-SCID with iPSC line source name designation in parenthesis, and where all subject blood cell sources are unique, with one exception being that the same blood sample from a single healthy subject iHC-04 was split to generate iPSC using STEMCCA-loxP (designated iHC-04) or using Sendai virus (designated iHC-04-s). (2) Donor age. (3) Whether MACS or Dynal was used for CD34+ cell purification. (4) Starting CD34+ cell number or blood volume as indicated. (5) Number of TRA-1-60 vital-stain positive iPSC colonies arising in the culture that were picked, expanded, and cryopreserved (for the Sendai reprogramming group, the estimated number arising in the culture is shown, but the number in parenthesis are the actual number of clones picked, successfully expanded, and cryopreserved). (6) Indicates whether “1” or “2” colonies from that group had completed verification of Cre-excision of single-copy STEMCCA-loxP insert, verification of source identity, continued expression of pluripotency markers, expansion, and cryopreservation (“0” indicates reprogramming source where Cre-excision of a clone with full-line qualification is still in progress). For the Sendai group, the issue of whether Cre-excision was done is not applicable. Note that identity testing for the Cre-excised lines from the 3 X-SCID patients included demonstration that the patient’s known mutation of IL2RG is present in the iPSC line. (7) This column indicates if “1” or “2” of these lines has fully completed either embryoid body three germ layer histology analysis and/or teratoma formation testing in NSG mice (“0” indicates this testing is still in progress); for the STEMCCA-loxP group, all those indicated in column 7 are fully characterized Cre-excised lines.
AR-CGD, autosomal recessive p22phox (due to CYBA mutation), p47phox (due to NCF1 mutation), or p67phox (due to NCF2 mutation) deficient CGD; Dynal, Dynal anti-CD34 magnetic immunobead system; MACS, Miltenyi anti-CD34 magnetic immunobead system; n/a, not applicable; PBMC, peripheral blood mononuclear cells; X-CGD, X-linked gp91phox deficient chronic granulomatous disease (due to CYBB mutation); X-SCID, X-linked severe combined immune deficiency (due to IL2RG mutation).