A5032471166- Corneal Surgery and Treatments
- Retinal Development and Disorders
- CRISPR and Genetic Engineering
- Ocular Surface and Contact Lens
- Corneal surgery and disorders
- Pluripotent Stem Cells Research
- Ocular Disorders and Treatments
- RNA and protein synthesis mechanisms
- Advanced biosensing and bioanalysis techniques
- Retinal and Macular Surgery
- Virus-based gene therapy research
- Neuroscience and Neural Engineering
- Biomedical Ethics and Regulation
- Retinal Diseases and Treatments
- Tissue Engineering and Regenerative Medicine
- RNA Research and Splicing
- Nuclear Structure and Function
- Genetics and Neurodevelopmental Disorders
- Neuroethics, Human Enhancement, Biomedical Innovations
- Mosquito-borne diseases and control
- Congenital limb and hand anomalies
- Ocular Oncology and Treatments
- Genomics and Chromatin Dynamics
- Liver physiology and pathology
- Oral and Maxillofacial Pathology
L V Prasad Eye Institute
2015-2024
Brien Holden Vision Institute
2021
Centre for Cellular and Molecular Biology
2005-2007
Corneal epithelial stem cells residing within the annular limbal crypts regulate adult tissue homeostasis. Autologous grafts and engineered corneal cell sheets have been widely used in treatment of various ocular surface defects. In case bilateral defects, pluripotent (PSC) derived are now being explored as an alternative to allogeneic grafts. We report here efficient method generate complex three dimensional organoids from human PSCs. The eye field primordial (EFP) clusters that emerged...
Abstract The clinical success of CRISPR therapies hinges on the safety and efficacy Cas proteins. Cas9 from Francisella novicida (FnCas9) is highly precise, with a negligible affinity for mismatched substrates, but its low cellular targeting efficiency limits therapeutic use. Here, we rationally engineer protein to develop enhanced FnCas9 (enFnCas9) variants broaden their accessibility across human genomic sites by ~3.5-fold. enFnCas9 proteins single mismatch specificity expanded target...
Stargardt retinopathy is an inherited form of macular degeneration caused by mutations in gene ABCA4 and characterized the accumulation lipid-rich deposits retinal pigment epithelium (RPE), RPE atrophy, photoreceptor cell death. Inadequate mechanistic insights into pathophysiological changes occurring have hindered disease treatments. Here, we show that knockout induced pluripotent stem cell-derived (STGD1-iRPE) from patients with differentiate normally but display intracellular lipid...
Leber congenital amaurosis, Type 12 is an early onset, autosomal recessive retinal disease caused by mutations in RD3. We report the generation of a patient-specific iPSC line (LVPEIi006-B), using Sendai viral vector-based reprogramming approach and isogenic, mutation-corrected (LVPEIi006-B-1), en31FnCas9-based adenine base editor (ABE) system. Both lines were clonally expanded genotyped to confirm presence mutation desired correction edited line. maintained their stemness, pluripotency,...
In Brief Purpose: To establish primary cultures of human nasal mucosal fibroblasts (HNMFs) and to test the effect varying concentrations mitomycin C (MMC) treatment durations on cellular proliferation viability fibroblasts. Design: Laboratory investigation. Methods: Nasal mucosa harvested from patients undergoing a dacryocystorhinostomy was used by explant culture method. Cells were expanded frozen at every passage, passage 3 cells for further experiments. The then treated with different...
Extensive damage to the limbal region of cornea leads a severe form corneal blindness termed as stem cell deficiency (LSCD). Whereas most cases opacity can be treated with full thickness transplants, LSCD requires transplantation for successful ocular surface reconstruction. Current treatments using involve use murine NIH 3T3 cells and human amniotic membranes culture substrates, which pose threat transmission animal-derived pathogens donor tissue-derived cryptic infections. In this study,...
To understand the response of oral epithelial cells, transplanted on corneal surface to ocular cues in vivo. The button obtained after penetrating keratoplasty (PK) an eye a patient with total limbal stem cell deficiency (LSCD), previously treated cultured mucosal transplantation (COMET) was examined by immunohistochemistry for expression keratins, p63, p75, PAX6, Ki-67, CD31, and CD34. COMET followed optical-PK has improved visual acuity 20/40 rendered stable surface. excised tissue showed...
The A-type lamins that localize in nuclear domains termed lamin speckles are reorganized and antigenically masked specifically during myoblast differentiation. This rearrangement was observed to be linked the myogenic program as speckles, stained with monoclonal antibody (mAb) LA-2H10, were MyoD-transfected fibroblasts induced transdifferentiate muscle cells. In C2C12 myoblasts, early differentiation cyclin D3-expressing Ectopic D3 reorganization myoblasts but not other cell types....
Leber Congenital Amaurosis 2 is an early onset retinal dystrophy that occurs due to mutation in RPE65 gene. Here, we report the generation of two patient specific induced pluripotent stem cell lines harboring nonsense mutations exon 7 (c.646A > T) and 9 (c.992G A) gene, respectively, which leads premature translational termination formation defective protein. These were generated by reprogramming human dermal fibroblast cells using integration-free, episomal constructs expressing stemness...
With a recent amendment, India joined other countries that have removed the legislative barrier toward use of human-relevant methods in drug development. Here, global stakeholders weigh on urgent need to globally harmonize guidelines standardization microphysiological systems. We discuss possible framework for establishing scientific confidence and regulatory approval these methods.
Abstract Stem cells at the limbus mediate corneal epithelial regeneration and regulate normal tissue homeostasis. Ex vivo cultured limbal transplantations are being widely practiced in treatment of stem cell deficiency. In this report, we examined whether niche that nurture coexist ex cultures. We also compared inherent differences between explant suspension culture systems terms spatial distribution their effect on proliferation, migration, differentiation vitro. report content both was...
Retinoblastoma is a pediatric intraocular cancer caused by biallelic inactivation of RB1 gene in retinal progenitor cells. Here, we report the generation patient-specific induced pluripotent stem cell (iPSC) line (LVPEIi002-A) from patient diagnosed with retinoblastoma and showing familial inheritance nonsense mutation (c.1735C > T) within exon 18 one two alleles. This RB1+/- iPSC line, LVPEIi002-A was generated reprogramming peri-orbital fat tissue derived mesenchymal cells stably expanded...
Pluripotent stem cells can generate complex tissue organoids that are useful for in vitro disease modeling studies and developing regenerative therapies. This protocol describes a simpler, robust, stepwise method of generating retinal hybrid culture system consisting adherent monolayer cultures during the first 4 weeks differentiation till emergence distinct, self-organized eye field primordial clusters (EFPs). Further, doughnut-shaped, circular, translucent neuro-retinal islands within each...
Abstract Mice lacking C3G (RapGEF1), a ubiquitously expressed protein essential for neuronal differentiation, show multiple defects in brain development. Function of neurogenesis is poorly defined. Here, we identify specific expression novel isoform mice and humans. This has an insert the Crk-binding region, generating polypeptide 175 kDa, unlike previously known 140 kDa form all other tissues. In adult mouse brain, seen neurons, but was not detectable GFAP-positive cells. levels were high...
Mutations in ABCA4 gene leads to the most common form of an inherited retinal disease namely, Stargardt disease, type 1. Here, we report generation two different patient-specific induced pluripotent stem cell lines (LVPEIi007-B and LVPEIi008-B), carrying identical homozygous mutation, (c.6088C>T) within exon 44 gene. These were generated by reprogramming dermal fibroblasts, using integration-free, Sendai viral vectors. Both stably expanded expressed stemness pluripotency markers,...