Meyer, J. S. et al. Modeling early retinal development with human embryonic and induced pluripotent stem cells. Proc. Natl Acad. Sci. USA 106, 16698-16703

Stem Cell Research Program, Waisman Center, Departments of Anatomy and Neurology, University of Wisconsin-Madison, 1500 Highland Avenue, Madison, WI 53705, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 09/2009; 106(39):16698-703. DOI: 10.1073/pnas.0905245106
Source: PubMed


Human pluripotent stem cells have the potential to provide comprehensive model systems for the earliest stages of human ontogenesis. To serve in this capacity, these cells must undergo a targeted, stepwise differentiation process that follows a normal developmental timeline. Here we demonstrate the ability of both human embryonic stem cells (hESCs) and induced pluripotent stem (iPS) cells to meet these requirements for human retinogenesis. Upon differentiation, hESCs initially yielded a highly enriched population of early eye field cells. Thereafter, a subset of cells acquired features of advancing retinal differentiation in a sequence and time course that mimicked in vivo human retinal development. Application of this culture method to a human iPS cell line also generated retina-specific cell types at comparable times in vitro. Lastly, altering endogenous signaling during differentiation affected lineage-specific gene expression in a manner consistent with established mechanisms of early neural and retinal cell fate determination. These findings should aid in the investigation of the molecular events governing retinal specification from human pluripotent stem cells.


Available from: Su-Chun Zhang, Feb 08, 2015
  • Source
    • "Similarly, others have investigated the development of other neuronal cell types, including cerebellar Purkinje cells (Wang et al., 2015), retinal cells (Meyer et al., 2009) and motor neurons (Toma et al., 2015). More recently, hiPSC-derived neurons have been used to study specific aspects of cell biology, such as mitochondrial biogenesis (O'Brien et al., 2015) and neuromuscular junction development (Yoshida et al., 2015). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Over recent years tremendous progress has been made towards understanding the molecular and cellular mechanism by which estrogens exert enhancing effects on cognition, and how they act as a neuroprotective or neurotrophic agent in disease. Currently, much of this work has been carried out in animal models with only a limited number of studies using native human tissue or cells. Recent advances in stem cell technology now make it possible to reprogram somatic cells from humans into induced pluripotent stem cells (iPSCs), which can subsequently be differentiated in neurons of specific lineages. Importantly, the reprogramming of cells allows for the generation of iPSCs that retains the genetic "makeup" of the donor. Therefore, it is possible to generate iPSC-derived neurons from patients diagnosed with specific diseases, that harbor the complex genetic background associated with the disorder. Here, we review the iPSC technology and how its currently being used to model neural development and neurological diseases. Furthermore, we explore whether this cellular system could be used to understand the role of estrogens in human neurons, and present preliminary data in support of this. We further suggest that the use of iPSC technology offers a novel system in which to not only further understand estrogens' effects in human cells, but in which to investigate the mechanism by which estrogens are beneficial in disease. Developing a greater understanding of these mechanisms in native human cells will also aid in the development of safer and more effective estrogen-based therapeutics. Copyright © 2015. Published by Elsevier Inc.
    Hormones and Behavior 07/2015; 26. DOI:10.1016/j.yhbeh.2015.06.014 · 4.63 Impact Factor
  • Source
    • "Considering that the human photoreceptor require more time to mature in vivo, it is possible that the photoreceptor cells derived using this method have artificial intracellular microenvironments. In fact, there are several protocols for retinal differentiation that involve months of culture [11,12,36,37]. However, the method used in the present study required less time and constitutes an efficient strategy for creating cells that can be used to examine pathogenic genes and screen novel therapeutics, which can be then applied in industrial uses. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Background Retinitis pigmentosa (RP) is an inherited human retinal disorder that causes progressive photoreceptor cell loss, leading to severe vision impairment or blindness. However, no effective therapy has been established to date. Although genetic mutations have been identified, the available clinical data are not always sufficient to elucidate the roles of these mutations in disease pathogenesis, a situation that is partially due to differences in genetic backgrounds. Results We generated induced pluripotent stem cells (iPSCs) from an RP patient carrying a rhodopsin mutation (E181K). Using helper-dependent adenoviral vector (HDAdV) gene transfer, the mutation was corrected in the patient’s iPSCs and also introduced into control iPSCs. The cells were then subjected to retinal differentiation; the resulting rod photoreceptor cells were labeled with an Nrl promoter-driven enhanced green fluorescent protein (EGFP)-carrying adenovirus and purified using flow cytometry after 5 weeks of culture. Using this approach, we found a reduced survival rate in the photoreceptor cells with the E181K mutation, which was correlated with the increased expression of endoplasmic reticulum (ER) stress and apoptotic markers. The screening of therapeutic reagents showed that rapamycin, PP242, AICAR, NQDI-1, and salubrinal promoted the survival of the patient’s iPSC-derived photoreceptor cells, with a concomitant reduction in markers of ER stress and apoptosis. Additionally, autophagy markers were found to be correlated with ER stress, suggesting that autophagy was reduced by suppressing ER stress-induced apoptotic changes. Conclusion The use of RP patient-derived iPSCs combined with genome editing provided a versatile cellular system with which to define the roles of genetic mutations in isogenic iPSCs with or without mutation and also provided a system that can be used to explore candidate therapeutic approaches.
    Molecular Brain 06/2014; 7(1):45. DOI:10.1186/1756-6606-7-45 · 4.90 Impact Factor
  • Source
    • "3-D optic vesicle-like structures (OVs) have also been reported using human iPSCs (Meyer et al., 2009; Phillips et al., 2012) and ESCs (Boucherie et al., 2013; Meyer et al., 2011, 2009; Nakano et al., 2012). Building on an earlier study (Meyer et al., 2009), Meyer et al. (2011) showed in 2011 that human iPSCs and ESCs could generate neuroepithelial-like clusters of retinal progenitors with numerous characteristics of developing optic vesicles. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Human pluripotent stem cells have made a remarkable impact on science, technology and medicine by providing a potentially unlimited source of human cells for basic research and clinical applications. In recent years, knowledge gained from the study of human embryonic stem cells and mammalian somatic cell reprogramming has led to the routine production of human induced pluripotent stem cells (hiPSCs) in laboratories worldwide. hiPSCs show promise for use in transplantation, high throughput drug screening, "disease-in-a-dish" modeling, disease gene discovery, and gene therapy testing. This review will focus on the first application, beginning with a discussion of methods for producing retinal lineage cells that are lost in inherited and acquired forms of retinal degenerative disease. The selection of appropriate hiPSC-derived donor cell type(s) for transplantation will be discussed, as will the caveats and prerequisite steps to formulating a clinical Good Manufacturing Practice (cGMP) product for clinical trials.
    Experimental Eye Research 06/2014; 123. DOI:10.1016/j.exer.2013.12.001 · 2.71 Impact Factor
Show more