Table 1.
A summary of salient experimental features from recent stem cell-to-retina data.
| Cell source | Method | Culture Condition | Type of Cells Generated |
Factors Tested | Markers Detected | Time to Photoreceptors |
Time to RPE Pigmentation |
Functional Tests | Transplantation Tested? |
Comparison to hESC |
Reference |
|---|---|---|---|---|---|---|---|---|---|---|---|
| Human Fibroblasts |
Adherent Culture |
Matrigel | RPE | B27, N2, Noggin, DKK1, IGF1, bFGF+- Nicotineamide, Activin, SU5402, VIP |
Pmel17, nitf, tyrosinase, Tyrp2, PEDF, bestrophin |
N/A | 14 days | Phagocytosis | No | hiPSCs less efficient |
61 |
| Human Fibroblasts |
Adherent Culture |
PDL/ Laminin | RPE | Nicotineamide, Activin, NIC, SB431542 |
Mitf, OTX2, bestrophin, RLBPI |
N/A | 4 weeks | Phagocytosis, ion transport, membrane potential, polarized VEGF secretion |
No | N/A | 65 |
| Human Fibroblasts |
Adherent Culture |
Defined conditions, Matrigel or VN- PAS Synthemax |
RPE | Blebbistatin and removal of FGF |
Mitf, RPE65 | N/A | 25-30 days | Phagocytosis | Yes; assessed survival and phagocytosis |
Similar | 60 |
| Human RPE cells | Embryoid body |
Suspension then gelatin |
RPE | Removal of FGF | Bestrophin, ZO1, TIR, OTX2, Pax6 |
N/A | 2-3 weeks | Phagocytosis | No | N/A | 49 |
| Human Fibroblasts |
Adherent Culture |
Defined conditions, tested gelatin, Matrigel, laminin, collagen-I, -IV, elastin |
RPE | Removal of FGF | PEDF, Mitf, OTX2, PMel17, ZO1, bestrophin |
N/A | 5 weeks | phagocytosis (on hESCs) |
No | iPSCs pigmented at signifcantly lower frequencies |
51 |
| Human Fibroblasts |
Adherent Culture |
MEFs then gelatin |
RPE | Removal of FGF | RLBP, MERIK, Mitf, OTX2, PMEL17, RPECs, ZO1, PEDF, ATP1B1, bestrophin |
N/A | 4 week | In-vitro and in- vivo phagocytosis |
Yes, RCS rats | hiPSC-RPE rejected by host retina while hESC survived 10 weeks |
52 |
| Mouse and Human Fibroblasts |
Embryoid body |
Suspension then PDL/laminin, fibronectin |
RPE, photoreceptors |
DKK1, Lefty A, (RA and taurine for phoreceptor cultures) |
MITF, PAX6, RX, CRX, recoverin, RHO |
120 | 40 days | None | No | Similar | 57 |
| Human Fibroblasts |
Adherent Culture |
MEFs or Ks27s | RPE | Removal of FGF | Mitf, OTX2, tyrosinase, Typ1, SILVER, ZO1, BEST, and after 6 months RPE65 |
N/A | 20-30 days | Phagocytosis | No | Similar | 50 |
| Human Fibroblasts |
Embryoid body |
Suspension then PDL/laminin, fibronectin |
RPE, photoreceptors |
Small molecules CKI-7 and SB- 431542 (RA and taurine for photoreceptor cultures |
CRALBP, Mitf, RPE65, ZO1, CHX10, CRX, recoverin, RHO |
120-140 | 40 days | Phagocytosis of latex beads |
No | Similar | 58 |
| Human Fibroblasts |
Embryoid body |
Suspension then laminin |
RPE, photoreceptors |
Removal of FGF; N2, B27 |
Mitf, RPE65, ZO1, bestrophin, CHX10, CRX, recoverin, opsin |
80 | 35 days | None | No | Similar | 54 |
| Human Fibroblasts |
Adherent Culture |
MEFs then Matrigel |
RPE | Removal of FGF | RPE65,ZO1 | N/A | 3-4 weeks | Phagocytosis (only on hESCs) |
No | hESCs appear to resemble fetal RPE more than hiPSCs |
53 |
| Human Fibroblasts from RP, LCA, Usher syndrome and LHON patients |
Adherent Culture |
Feeder-free Matrigel |
RPE | Noggin, bFGF, RA, Shh |
MITF, RPE65, CRALBP, BEST, PAX6, OTX2, ZO1 |
N/A | 35 days | None | No | N/A | 56 |
| Fibroblasts of RP patients |
Embryoid body |
Suspension then PDL/ laminin/ fibronectin |
RPE, photoreceptors |
Lefty A, SB- 431542, DKK1, taurine, RA |
PAX6, RX, MITF, recoverin, rhodopsin |
60-110 days | 30 days | None | No | N/A | 68 |
| Fibroblasts from normal, RP, LCA, and Stargadt patients; also iris pigment epithelium |
Embryoid body |
Suspension then Synthemax for xeno-free and Matrigel for feeder-free |
RPE, photoreceptors, retinal ganglion cells |
B27, n2, noggin, DKK1, IGF1, DAPT, FGF |
PAX6, RX, MITF, ZO- 1, recoverin, rhodopsin, CHX10, NRL, BRN3b, NF200 |
90 Days | before 60 days |
None | No | N/A | 63 |
| Human Fibroblasts |
Adherent Culture |
Feeder-free Matrigel |
RPE, photoreceptors, retinal ganglion cells |
B27, N2, noggin, DKK1, IGF1 |
OTX2, Crx, Nrl, arrestin, recoverin, Trb2, rhodopsin and Pax6, ZO1, Hu C/D, BRN3, |
2 months | Not mentioned |
None | Yes, subretinal transplant of FACS sorted IRBP-GFP- expressing photoreceptors |
Similar | 70 |
| Human Fibroblasts |
Embryoid body |
Suspension or Matrigel for feeder-free and Synthemax for xeno-free cultures |
RPE, neural retina |
B27, N2, heparin (no heparin in xeno-free) |
PAX6, RX, MITF, CHX10, OTX2, BEST1, ZO1, PEDF, recoverin, CRX, BRN3 |
60 days | 1 month | None | No | Compared feeder, feeder-free and xeno- free; all similar results |
64 |