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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.