3D culture of human pluripotent stem cells in RGD-alginate hydrogel improves retinal tissue development.
No treatments exist to effectively treat many retinal diseases. Retinal pigmented epithelium (RPE) and neural retina can be generated from human embryonic stem cells/induced pluripotent stem cells (hESCs/hiPSCs). The efficacy of current protocols is, however, limited. It was hypothesised that generation of laminated neural retina and/or RPE from hiPSCs/hESCs could be enhanced by three dimensional (3D) culture in hydrogels. hiPSC- and hESC-derived embryoid bodies (EBs) were encapsulated in 0.5% RGD-alginate; 1% RGD-alginate; hyaluronic acid (HA) or HA/gelatin hydrogels and maintained until day 45. Compared with controls (no gel), 0.5% RGD-alginate increased: the percentage of EBs with pigmented RPE foci; the percentage EBs with optic vesicles (OVs) and pigmented RPE simultaneously; the area covered by RPE; frequency of RPE cells (CRALBP+); expression of RPE markers (TYR and RPE65) and the retinal ganglion cell marker, MATH5. Furthermore, 0.5% RGD-alginate hydrogel encapsulation did not adversely affect the expression of other neural retina markers (PROX1, CRX, RCVRN, AP2α or VSX2) as determined by qRT-PCR, or the percentage of VSX2 positive cells as determined by flow cytometry. 1% RGD-alginate increased the percentage of EBs with OVs and/or RPE, but did not significantly influence any other measures of retinal differentiation. HA-based hydrogels had no significant effect on retinal tissue development. The results indicated that derivation of retinal tissue from hESCs/hiPSCs can be enhanced by culture in 0.5% RGD-alginate hydrogel. This RGD-alginate scaffold may be useful for derivation, transport and transplantation of neural retina and RPE, and may also enhance formation of other pigmented, neural or epithelial tissue.



The burden of retinal disease is ever growing with the increasing age of the world-wide population. Transplantation of retinal tissue derived from human pluripotent stem cells (PSCs) is considered a promising treatment. However, derivation of retinal tissue from PSCs using defined media is a lengthy process and often variable between different cell lines. This study indicated that alginate hydrogels enhanced retinal tissue development from PSCs, whereas hyaluronic acid-based hydrogels did not. This is the first study to show that 3D culture with a biomaterial scaffold can improve retinal tissue derivation from PSCs. These findings indicate potential for the clinical application of alginate hydrogels for the derivation and subsequent transplantation retinal tissue. This work may also have implications for the derivation of other pigmented, neural or epithelial tissue.
Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle NE1 3BZ, UK. Electronic address: nicola.hunt@ncl.ac.uk.
Nicola C
NC
Hunt
Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle NE1 3BZ, UK. Electronic address: dean.hallam@ncl.ac.uk.
Dean
D
Hallam
Cumberland Infirmary, North Cumbria University Hospitals NHS Trust, Carlisle CA2 7HY, UK.
Ayesha
A
Karimi
Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle NE1 3BZ, UK. Electronic address: carla.mellough@ncl.ac.uk.
Carla B
CB
Mellough
School of Mechanical & Systems Engineering, Stephenson Building, Newcastle University, Newcastle upon Tyne, UK. Electronic address: jinju.chen@ncl.ac.uk.
Jinju
J
Chen
Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle NE1 3BZ, UK; Sunderland Eye Infirmary, Queen Alexandra Road, Sunderland SR2 9HP, UK. Electronic address: david.steel@ncl.ac.uk.
David H W
DHW
Steel
Institute of Genetic Medicine, Newcastle University, International Centre for Life, Central Parkway, Newcastle NE1 3BZ, UK. Electronic address: majlinda.lako@ncl.ac.uk.
Majlinda
M
Lako
D000464:Alginates / Q000494:pharmacology*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D020723:Glucuronic Acid / Q000494:pharmacology; D006603:Hexuronic Acids / Q000494:pharmacology; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000494:pharmacology*; D009842:Oligopeptides / Q000494:pharmacology*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D012160:Retina / Q000187:drug effects / Q000254:growth & development*; D055213:Retinal Pigment Epithelium / Q000166:cytology
D000464:Alginates; D006603:Hexuronic Acids; D009842:Oligopeptides; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C047981:arginyl-glycyl-aspartic acid; D020723:Glucuronic Acid
Biomaterials; Embryonic stem cells; Induced pluripotent stem cells; Retina; Tissue engineering