Table 1 Derivation of LESCs and CECs from hiPSCs
From: Corneal cell therapy: with iPSCs, it is no more a far-sight
Authors/year/reference number | Stem cell type | Cell type | Time line in days | Culture conditions | Remarks |
|---|---|---|---|---|---|
Ahmad et al./2007/40 | ESC | CEP | 5 | Differentiated on collagen IV (ColIV)/laminin/fibronectin-coated substrate and limbal fibroblast-conditioned medium | ColIV was found to be a better substrate compared to laminin and fibronectin. |
Hayashi et al./2012/24 | iPSC | CEP | 100 | Differentiated on gelatin-coated substrate and the latter on PA6 feeder layer (stromal-derived inducing activity) in GMEM culture medium + 10% KOSR | Human CEPs were reprogrammed to iPSC using lentivirus. iPSCs were cultured in feeder-dependent conditions. |
Brzeszczynska et al./2014/42 | ESC | CEP | 21 | Differentiated on Matrigel substrate in human limbal fibroblast-conditioned medium | Derived CEPs were characterized for P63, ABCG2, and CK expression. |
Michailova et al./2014/46 | iPSC | CEP | 44 | D1–4 suspension culture: Cnt-30 medium supplemented with TGFβ and WNT inhibitors or in basal RegES medium. D5–44 adherent cultures on collagen IV-coated substrates in CnT-30 medium supplemented with TGFβ and WNT inhibitors | Derived CEPs were characterized for P63, PAX6, CK3, CK12, and CK15. |
Michailova et al./2015/38 | ESC/iPSC | LESC | 35 | Suspension culture: cultured in ESC medium + inhibitors of TGFβ and WNT Adherent culture: cultured on ColIV-coated substrates in CnT-30 medium | Comparative proteomics reveal iPSC-derived LESCs are similar to native ocular surface epithelial cells |
Hayashi et al./2016/114 | iPSC | CEP | 100 | D0–28: on laminin substrate in differentiation medium: GMEM + KOSR + NEAA + Na-pyruvate D29–56: on laminin substrate in corneal differentiation medium | The authors mention the use of an appropriate iPSC clone is import to achieve CEP differentiation |
D57–70: CnT – PR: DMEM + keratocyte growth factor + RI D71–100: on laminin substrate in corneal epithelial maintenance medium | |||||
Cieślar-Pobuda et al./2016/45 | iPSC | CEP | 21 | Cultured on gelatin-coated substrate in human limbal fibroblast-conditioned medium | hDF is reprogrammed to iPSC using lentivirus. Derived characterized for P63, ABCG2, PAX6, and cytokeratin expression. |
Aberdam et al./2017/47 | iPSC | LESC | 35 | D0–4: induction medium with TGFβ inhibitor + BMP4* | Modified the culture condition to produce a propagatable pure population of iPSC-derived LEC (LiPSC). *Hayashi et al. [19] show BMP4 treatment suppressed CEP differentiation from iPSCs. |
D5–21: cultured as monolayer on collagen IV substrate in induction medium with TGFβ inhibitor + BMP4 + EGF + RI | |||||
D22–35: Induction medium with keratocyte growth factor + RI on collagen IV substrate. | |||||
Garcia de la Torre et al./2017/39 | iPSCs | LESCs | 14 | D0–1: EBs in complete essential medium 6 + inhibitors of TGFβ, WNT + bFGF D2–14: Corneal epithelial limbal-conditioned medium | Derived LESCs were validated for the expression of PAX6, P63, and cytokeratins. |
Zhang et al./2017/49 | ESCs | LESCs | 9 | D0–1: ES medium | Higher CO2 has beneficial effects on the differentiation of corneal epithelial progenitor cells (CEPCs) from hESCs. |
D2–9: cultured under 5%/7%/9% CO2 | |||||
In keratocyte serum-free medium + DMEM/F12 on ColIV substrate | |||||
Kamarudin et al./2017/43 | ESCs/iPSCs | CEP | 20 | D0–1: mTesR medium + RI D2–20: compared different composition of differentiation medium with TGFβ, WNT inhibitors. On D9, cultures were plated on collagen IV-coated substrate | A two-step protocol reporting better CEP differentiation efficacy. The work [35] reveals a differential ability of hiPSC lines to generate CEPs underlined by the activity of endogenous BMP signaling pathway. |
Hongsito et al./2017/44 | ESCs/iPSCs | LESCs | 143 | D0–1: suspension culture; inhibition of TGFβ and WNT pathway; addition of bFGF and BMP-4 D2–143: monolayer culture on laminin and collagen IV-coated substrates in CNT-30 medium | Methodology to produce two clinically relevant ocular epithelial cell types from feeder- and xeno-free hPSC. |