A5090913554- Pluripotent Stem Cells Research
- CRISPR and Genetic Engineering
- Connective tissue disorders research
- Renal and related cancers
- 3D Printing in Biomedical Research
- Peptidase Inhibition and Analysis
- Genomic variations and chromosomal abnormalities
- Tissue Engineering and Regenerative Medicine
- Animal Genetics and Reproduction
- Reproductive Biology and Fertility
- Dermatological and Skeletal Disorders
- RNA Interference and Gene Delivery
- Bone and Dental Protein Studies
- Biomedical Ethics and Regulation
- Genetics and Neurodevelopmental Disorders
- Genomics and Chromatin Dynamics
- Chromosomal and Genetic Variations
- Bone health and treatments
- Reproductive System and Pregnancy
- Wnt/β-catenin signaling in development and cancer
- Dupuytren's Contracture and Treatments
- Cancer Genomics and Diagnostics
- PI3K/AKT/mTOR signaling in cancer
- Gene Regulatory Network Analysis
- Pregnancy and preeclampsia studies
Sheffield Children's Hospital
2008-2025
Sheffield Children's NHS Foundation Trust
2012-2025
University of Sheffield
2009-2023
National Health Service
2015
Boston Children's Hospital
2010-2011
Boston Children's Museum
2011
Children’s Foundation
2010
Much attention has focussed on the conversion of human pluripotent stem cells (PSCs) to a more naïve developmental status. Here we provide method for resetting via transient histone deacetylase inhibition. The protocol is effective across multiple PSC lines and can proceed without karyotype change. Reset be expanded feeders with doubling time around 24 h. WNT inhibition stabilises process. transcriptome reset diverges markedly from that primed PSCs shares features inner cell mass (ICM)....
Abstract Lack of robust methods for establishment and expansion pluripotent human embryonic stem (hES) cells still hampers development cell therapy. Laminins (LN) are a family highly cell-type specific basement membrane proteins important adhesion, differentiation, migration phenotype stability. Here we produce isolate recombinant LN-521 isoform develop culture matrix containing E-cadherin, which both localize to niches in vivo . This allows clonal derivation, survival long-term self-renewal...
A major limitation in developing applications for the use of human embryonic stem cells (HESCs) is our lack knowledge their responses to specific cues that control self-renewal, differentiation, and lineage selection. HESCs are most commonly maintained on inactivated mouse fibroblast feeders medium supplemented with FCS, or proprietary replacements such as knockout serum-replacement together FGF-2. These undefined culture conditions hamper analysis mechanisms HESC behavior. We have now...
Human embryonic stem cells (hESCs) regularly acquire nonrandom genomic aberrations during culture, raising concerns about their safe therapeutic application.The International Stem Cell Initiative identified a copy number variant (CNV) amplification of chromosome 20q11.21 in 25% hESC lines displaying normal karyotype.By comparing four cell paired for the presence or absence this CNV, we show that those containing amplicon have higher population doubling rates, attributable to enhanced...
Genetic changes in human pluripotent stem cells (hPSCs) gained during culture can confound experimental results and potentially jeopardize the outcome of clinical therapies. Particularly common hPSCs are trisomies chromosomes 1, 12, 17, 20. Thus, should be regularly screened for such aberrations. Although a number methods used to assess hPSC genotypes, there has been no systematic evaluation sensitivity commonly techniques detecting low-level mosaicism cultures. We have performed mixing...
In contrast to conventional human pluripotent stem cells (hPSCs) that are related post-implantation embryo stages, naive hPSCs exhibit features of pre-implantation epiblast. Naive established by resetting hPSCs, or derived from dissociated inner cell masses. Here we investigate conditions for transgene-free reprogramming somatic pluripotency. We find Wnt inhibition promotes RNA-mediated induction demonstrate application independent fibroblast cultures and endothelial progenitor cells. show...
Human embryonic stem cells (hESCs) have potential use in clinical therapy and regenerative medicine. One of the major challenges regarding application these is development an efficient cryopreservation protocol, since current methods, which include slow-freezing-rapid thawing vitrification colonies suspension, present poor viability high differentiation rates. Dissociated hESC suspensions do not survive because they are susceptible to apoptosis upon cell detachment dissociation. A selective...
Both human embryonic stem cells (hESCs) and induced pluripotent (iPSCs) bear a great potential in regenerative medicine. In addition to optimized clinical grade culture conditions, efficient cryopreservation methods for these are needed. Obtaining good survival after thawing has been problematic. We used novel, chemically defined effective xeno-free system cryostorage banking of hESCs iPSCs. The earlier established slow freezing protocols have, even recent improvements, resulted low...
Chromosome 20 abnormalities are some of the most frequent genomic changes acquired by human pluripotent stem cell (hPSC) cultures worldwide. Yet their effects on differentiation remain largely unexplored. We investigated a recurrent abnormality also found amniocentesis, isochromosome 20q (iso20q), during clinical retinal pigment epithelium differentiation. Here we show that iso20q interrupts spontaneous embryonic lineage specification. Isogenic lines revealed under conditions promote...
Culture-acquired variants in human pluripotent stem cells (hPSCs) hinder their applications research and clinic. However, the mechanisms that underpin selection of remain unclear. Here, through analysis comprehensive karyotyping datasets from over 23,000 hPSC cultures more than 1,500 lines, we explored how culture conditions shape variant selection. Strikingly, identified an association chromosome 1q gains with feeder-free noted a rise its prevalence recent years, coinciding increased usage...
Abstract Background Cell therapies based on human pluripotent stem cells (hPSCs) are in clinical trials with the aim of restoring vision people age-related macular degeneration. The final cell therapy product consists retinal pigment epithelium (RPE) differentiated from hPSCs. However, hPSCs recurrently acquire genetic abnormalities that give them an advantage culture unknown effects to clinically-relevant progeny. One most common is sub-karyotype 20q11.21 copy number variant, known carry...
Abstract Hypophosphatasia (HPP) is an inborn error of metabolism caused by loss-of-function variants in the ALPL gene, which encodes tissue non-specific isozyme alkaline phosphatase (ALP). There no typical phenotype adults. We used a genotyping first approach to determine whether pathogenic were associated with musculoskeletal symptoms, mineral abnormalities, and impact on quality life. recruited individuals (or likely pathogenic) variant gene (n = 26) their relatives 44). performed genetic...
Summary Teratocarcinomas are a subset of tumours that result from the neoplastic transformation primordial germ cells. Such cell (GCT) histologically heterogeneous, reflecting capacity for differentiation (pluripotency) their embryonal carcinoma (EC) stem However, malignant evolution these may ultimately correlate with decrease in pluripotency, because this would tend to increase propensity EC cells self‐renewal. Human embryonic (ES) cells, derived early blastocysts, closely resemble and, on...
Genomic integrity of human pluripotent stem cell (hPSC) lines requires routine monitoring. We report here that novel karyotyping assay, utilizing bead-bound bacterial artificial chromosome probes, provides a fast and easy tool for detection chromosomal abnormalities in hPSC lines. The analysis can be performed from low amounts DNA isolated whole pools with simple data interface. method enables screening cost-efficient high-throughput manner.
Background The Ehlers-Danlos syndromes (EDS) consist of 13 subtypes with overlapping features including joint hypermobility, skin and vascular fragility generalized connective tissue friability. As DNA analysis has become the gold standard for investigation EDS, transmission electron microscopy (TEM) in clinical practice is decreasing. However, owing to use next-generation sequencing, frequency variants uncertain significance (VUS) identified using increasing. We hypothesized that TEM can...