A5063963894- Retinal Development and Disorders
- CRISPR and Genetic Engineering
- Photoreceptor and optogenetics research
- Neuroscience and Neural Engineering
- Pluripotent Stem Cells Research
- Photochromic and Fluorescence Chemistry
- Retinal Diseases and Treatments
- 3D Printing in Biomedical Research
- bioluminescence and chemiluminescence research
- Ocular Disorders and Treatments
- Virus-based gene therapy research
- RNA regulation and disease
- Neurogenesis and neuroplasticity mechanisms
- Ocular Oncology and Treatments
- Single-cell and spatial transcriptomics
- melanin and skin pigmentation
- Corneal Surgery and Treatments
- Retinopathy of Prematurity Studies
- Ocular and Laser Science Research
- Protein Kinase Regulation and GTPase Signaling
- Corneal surgery and disorders
- Nitric Oxide and Endothelin Effects
- Glaucoma and retinal disorders
- Genomics and Rare Diseases
- Retinal and Macular Surgery
University of Wisconsin–Madison
2016-2025
Smith-Kettlewell Eye Research Institute
2016-2025
McPherson College
2017-2023
Visual Sciences (United States)
2014-2021
University of Wisconsin System
2021
Pisgah Astronomical Research Institute
2008-2017
University of Iowa Hospitals and Clinics
2005
Retina Foundation of the Southwest
2005
Johns Hopkins University
2005
The University of Texas Southwestern Medical Center
2005
Human pluripotent stem cells have the potential to provide comprehensive model systems for earliest stages of human ontogenesis. To serve in this capacity, these must undergo a targeted, stepwise differentiation process that follows normal developmental timeline. Here we demonstrate ability both embryonic (hESCs) and induced (iPS) meet requirements retinogenesis. Upon differentiation, hESCs initially yielded highly enriched population early eye field cells. Thereafter, subset acquired...
Abstract Differentiation methods for human induced pluripotent stem cells (hiPSCs) typically yield progeny from multiple tissue lineages, limiting their use drug testing and autologous cell transplantation. In particular, early retina forebrain derivatives often intermingle in cultures, owing to shared ancestry tightly coupled development. Here, we demonstrate that three-dimensional populations of retinal progenitor (RPCs) can be isolated both embryonic hiPSC providing a valuable tool...
Numerous protocols have been described that produce neural retina from human pluripotent stem cells (hPSCs), many of which are based on the culture 3D organoids. While nearly all such methods yield at least partial segments highly mature-appearing retinal structure, variabilities exist within and between organoids can change over a protracted time course differentiation. Adding to this complexity potential differences in composition configuration when viewed across multiple differentiations...
Peutz-Jeghers syndrome (PJS) is an autosomal dominant disease characterized by melanocytic macules, hamartomatous polyps and increased risk for numerous cancers. The human LKB1 (hLKB1) gene encodes a serine/threonine protein kinase that deficient in the majority of patients with PJS. murine (mLKB1) cDNA was isolated, sequenced shown to produce 2.4-kb transcript encoding 436 amino acid 90% identity hLKB1. RNA blot RNase-protection analysis revealed mLKB1 mRNA expressed all tissues cell lines...
Best disease (BD) is an inherited degenerative of the human macula that results in progressive and irreversible central vision loss. It caused by mutations retinal pigment epithelium (RPE) gene BESTROPHIN1 (BEST1), which, through mechanism(s) remain unclear, lead to accumulation subretinal fluid autofluorescent waste products from shed photoreceptor outer segments (POSs). We employed iPS cell (hiPSC) technology generate RPE BD patients unaffected siblings order examine cellular molecular...
Purpose.: We sought to determine if human induced pluripotent stem cells (iPSCs) derived from blood could produce optic vesicle–like structures (OVs) with the capacity stratify and express markers of intercellular communication. Methods.: Activated T-lymphocytes a routine peripheral sample were reprogrammed by retroviral transduction iPSCs. The T-lymphocyte–derived iPSCs (TiPSCs) characterized for pluripotency differentiated OVs using our previously published protocol. TiPSC-OVs then...
Gene-corrected patient-specific induced pluripotent stem (iPS) cells offer a unique approach to gene therapy. Here, we begin assess whether the mutational load acquired during correction of iPS is compatible with use in treatment genetic causes retinal degenerative disease. We isolated free transgene sequences from patient gyrate atrophy caused by point mutation encoding ornithine-δ-aminotransferase ( OAT ) and used homologous recombination correct defect. Cytogenetic analysis, array...
To determine the effects of serial expansion on cellular, molecular, and functional properties human iPS cell (hiPSC)-derived RPE cultures.Fibroblasts obtained from four individuals were reprogrammed into hiPSCs differentiated to cells using previously described methods. Patches deeply pigmented hiPSC-RPE dissected, dissociated, grown in culture until they re-formed monolayers. Subsequent passages by repeated dissociation, expansion, maturation Gene protein expression profiles morphological...
Human induced pluripotent stem cells (hiPSCs) have been shown to differentiate along the retinal lineage in a manner that mimics normal mammalian development. Under certain culture conditions, hiPSCs form optic vesicle-like structures (OVs), which contain proliferating progenitors capable of yielding all neural retina (NR) cell types over time. Such observations imply conserved roles for regulators retinogenesis hiPSC-derived cultures and developing embryo. However, whether what extent this...
Significance Age-related macular degeneration (AMD) and related dystrophies (MDs) are a major cause of vision loss. However, pharmacological treatments in these diseases limited due to the lack knowledge underlying disease mechanisms, partly because appropriate human models study AMD MDs lacking. Furthermore, living eye, entire retina acts as functional unit, making it difficult investigate specific contribution particular retinal cell type disease. Here, we established multiple MDs, which...
Highlights•Unique transcriptional profiles demonstrate diversity among hPSC-derived RGCs•Numerous RGC subtypes characterized from RGCs•Molecular markers identified for through single-cell RNA-seq analysisSummaryRetinal ganglion cells (RGCs) are the projection neurons of retina and transmit visual information to postsynaptic targets in brain. While this function is shared nearly all RGCs, class cell remarkably diverse, comprised multiple subtypes. Previous efforts have numerous animal models,...
Abstract Photoreceptor loss is a leading cause of blindness, but mechanisms underlying photoreceptor degeneration are not well understood. Treatment strategies would benefit from improved understanding gene-expression patterns directing development, as many genes implicated in both development and degeneration. Neural retina leucine zipper (NRL) critical for rod genesis degeneration, with NRL mutations known to enhanced S-cone syndrome retinitis pigmentosa. While murine Nrl has been...
BackgroundA promising clinical application for stem and progenitor cell transplantation is in rescue therapy degenerative diseases. This strategy seeks to preserve rather than restore host tissue function by taking advantage of unique properties often displayed these versatile cells. In studies using different neurodegenerative disease models, transplanted human neural cells (hNPC) protected dying neurons within both the brain spinal cord. Based on reports, we explored potential hNPC visual...
As a follow-up to previous studies showing that human cortical neural progenitor cells (hNPC(ctx)) can sustain vision for at least 70 days after injection into the subretinal space of Royal College Surgeons (RCS) rats, authors examined how functional rescue is preserved over long periods and this relates retinal integrity donor cell survival.Pigmented dystrophic RCS rats (n = 15) received unilateral injections hNPC(ctx) postnatal day (P) 21; control 10) medium alone were untreated. All...
Abstract The protocol outlined below is used to differentiate human pluripotent stem cells (hPSCs) into retinal cell types through a process that faithfully recapitulates the stepwise progression observed in vivo. From pluripotency, are differentiated primitive anterior neural fate, followed by two distinct populations of progenitors and forebrain progenitors, each which can be manually separated purified. hPSC‐derived found self‐organize three‐dimensional optic vesicle−like structures, with...
Loss of photoreceptor cells due to retinal degeneration is one the main causes blindness in developed world. Although there currently no effective treatment, cell replacement therapy using stem-cell-derived may be a feasible future treatment option. In order ensure safety and efficacy this approach, robust isolation purification protocols must developed. To end, we previously biomarker panel for mouse precursors from developing retina embryonic stem cultures. current study applied approach...
Purpose: To report the clinical course of 6 patients with refractory neurotrophic corneal ulcers that were treated topical insulin drops. Methods: Retrospective chart review who had or epithelial defects to standard medical and surgical treatment. Insulin drops, prepared by mixing regular in artificial tears a polyethylene glycol propylene base at concentration 1 unit per milliliter, prescribed 2 3 times daily. Results: Six patients, aged 73 years, developed range treatments, including...
Genome-edited human pluripotent stem cells (hPSCs) have broad applications in disease modeling, drug discovery, and regenerative medicine. We present characterize a robust method for rapid, scarless introduction or correction of disease-associated variants hPSCs using CRISPR/Cas9. Utilizing non-integrated plasmid vectors that express puromycin N-acetyl-transferase (PAC) gene, whose expression translation is linked to Cas9, we transiently select based on their early levels Cas9 protein. Under...