Accelerated Three-Dimensional Neuroepithelium Formation from Human Embryonic Stem Cells and Its Use for Quantitative Differentiation to Human Retinal Pigment Epithelium

Abstract

Successful applications of pluripotent stem cells to cell-based therapies will rely on rapid and efficient methods to differentiate cells toward the target cell type. While methods have been developed for the generation of some medically relevant cell types including retinal pigment epithelium (RPE) cells, such protocols are lengthy and result in a heterogeneous cell mixture of RPE and non-RPE cells, requiring manual subselection and expansion. Such considerations have significant limiting impact of therapeutic applicability. Here we describe the accelerated three-dimensional neuroepithelial cyst culture of human embryonic stem cells (hESCs) and its utility to achieve quantitative production of RPE cell sheet with no manual selection in 30 days.

Salamanders: The molecular basis of tissue regeneration and its relevance to human disease

“…In this chapter, the regeneration of some of the tissues and organs in salamanders was included, these recent advances show the potential to translate this knowledge to develop new therapies in humans. For example; in the case of the eye, the conservation of the cell types and genetic pathways illustrate the potential for this knowledge to be used to design novel translational therapies (Barbosa-Sabanero et al, 2012;Carido et al, 2014;Chiba, 2014;Del Rio-Tsonis et al, 1998;Hayashi et al, 2004Hayashi et al, , 2013Haynes, Gutierrez, Aycinena, Tsonis, & Del Rio-Tsonis, 2007;Islam et al, 2014;Zhu et al, 2013;Zhu, Schreiter, & Tanaka, 2016). Similarly, the research on spinal cord regeneration in salamanders has identified critical molecular pathways that are conserved in mammals but which axolotls specifically activate to promote regeneration instead of glial scar formation (Diaz Quiroz, Tsai, Coyle, Sehm, & Echeverri, 2014;Sabin et al, 2019Sabin et al, , 2015, this knowledge of the axolotl spinal cord may help to inform potential cell based therapeutics in the future (Albors et al, 2015;Diaz Quiroz, Li, Aparicio, & Echeverri, 2016;Meinhardt et al, 2014).…”

Transplantable and functional retinal pigment epithelium derived from non-colony-type monolayer of human induced pluripotent stem cells

“…In 2004, Klimanskaya et al first reported that putative RPE cells could be derived from spontaneous differentiation of ESCs [12]. Many other studies about RPE cells derived from ESCs can also be found [13][14][15]. However, the application of ESCs has ethical difficulties as well as immune rejection problems.…”

Progress of stem/progenitor cell-based therapy for retinal degeneration

“…However, these limitless possibilities of ESCs have also led to ethical concerns, and there are difficulties in limiting the differentiation of ESCs into ideal photoreceptor cells in vitro and in vivo [66]. As a result, protocols are being established to conform to Good Manufacturing Practices, which include (1) the regulation of micro-environment to obtain a relatively high yield of target cells [67]; (2) manipulation of signalling pathways (such as Rho-associated, coiled-coil protein kinase inhibitions) to help eye formation and enhance hESC expansion [68]; (3) feeder-free strategies to improve differentiation efficiency [69]; (4) xeno-free techniques (free of human or animal derivatives) to greatly reduce the risk of contamination in harvested cells and decrease immune response [70]; (5) three-dimensional retina cultures intended to form complete and organized retinas to better mediate retinal repair [71, 72]; 6) bioengineering techniques, such as porous honeycomb-like films [73] and ultrathin substrates [74] to ensure high adherence and differentiation of hESC-RPE before and after implantation.…”