ArticleTissue engineering. Part C, Methods2011
A new isolation method of human limbal progenitor cells by maintaining close association with their niche cells.
Article in Tissue engineering. Part C, Methods, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 76 papers.
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Who cites it
76 citing papers in PubMed, 136 citations in OpenAlex.
- Constitutive expression of pentraxin 3 (PTX3) protein by human amniotic membrane cells leads to formation of the heavy chain (HC)-hyaluronan (HA)-PTX3 complex.The Journal of biological chemistry · 2014Trial
- Preclinical Models of Rare Corneal and Ocular Surface Diseases: a Comprehensive Narrative Review.Ophthalmology and therapy · 2026Review
- Progenitor cells from limbal, conjunctival and oral mucosal biopsies cultivated on silicone hydrogel contact lenses share CD90, CK15, nestin, CXCR4 and SDF-1 niche markers but preserve tissue-specific characteristics and differentiation directions.Frontiers in cell and developmental biology · 2026Article
- Reprogramming of Human Limbal Niche Progenitors.International journal of medical sciences · 2026Review
- Proinflammatory gene and protein expression alterations in human limbal aniridia fibroblasts.PloS one · 2025Article
- Anti-Inflammatory and Anti-(Lymph)angiogenic Properties of an ABCB5+ Limbal Mesenchymal Stem Cell Population.International journal of molecular sciences · 2024Article
- [Regenerative medicine for the corneal epithelium : Cell therapy from bench to bedside].Die Ophthalmologie · 2022Review
- Transcriptomic Profiling of Human Limbus-Derived Stromal/Mesenchymal Stem Cells-Novel Mechanistic Insights into the Pathways Involved in Corneal Wound Healing.International journal of molecular sciences · 2022Article
- Isolation andBio-protocol · 2022Article
- Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus.International journal of molecular sciences · 2022Article
- Article
- Human limbal epithelial stem cell regulation, bioengineering and function.Progress in retinal and eye research · 2021Review
- Long-term survival of cultivated oral mucosal epithelial cells in human cornea: generating cell sheets using an animal product-free culture protocol.Stem cell research & therapy · 2021Article
- Stem Cell Niche Microenvironment: Review.Bioengineering (Basel, Switzerland) · 2021Review
- Process development and safety evaluation of ABCB5Stem cell research & therapy · 2021Article
- HC-HA/PTX3 from amniotic membrane reverts senescent limbal niche cells to Pax6+ neural crest progenitors to support limbal epithelial progenitors.Stem cells (Dayton, Ohio) · 2021Article
- Corneal stromal mesenchymal stem cells: reconstructing a bioactive cornea and repairing the corneal limbus and stromal microenvironment.International journal of ophthalmology · 2021Review
- A Comparison of Methods for Isolation of Limbal Niche Cells: Maintenance of Limbal Epithelial Stem/Progenitor Cells.Investigative ophthalmology & visual science · 2020Article
- Isolation and enrichment of melanocytes from human corneal limbus using CD117 (c-Kit) as selection marker.Scientific reports · 2020Article
- Niche regulation of limbal epithelial stem cells: HC-HA/PTX3 as surrogate matrix niche.Experimental eye research · 2020Review
16 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 3 countries.
Funding
Abstract
In human corneal epithelium, self-renewal and fate decision of stem cells are highly regulated in a niche microenvironment called palisades of Vogt in the limbus. Herein, we discovered that digestion with dispase, which cleaves off the basement membrane, did not remove the entire basal epithelial progenitor cells. In contrast, digestion with collagenase isolated on cluster consisting of not only entire epithelial progenitor cells but also their closely associated mesenchymal cells because of better preservation of some basement membrane matrix. Collagenase isolated more basal epithelial progenitor cells, which were p63α+ and small in the size (8 μm in diameter), and generated significantly more holoclones and meroclones on 3T3 fibroblast feeder layers than dispase. Further, collagenase isolated more small pan-cytokeratin-/p63α-/vimentin+ cells with the size as small as 5 μm in diameter and heterogeneously expressing vimentin, Oct4, Sox2, Nanog, Rex1, Nestin, N-cadherin, SSEA4, and CD34. Maintenance of close association between them led to clonal growth in a serum-free, low-calcium medium, whereas disruption of such association by trypsin/EDTA resulted in no clonal growth unless cocultured with 3T3 fibroblast feeder layers. Similarly, on epithelially denuded amniotic membrane, maintenance of such association led to consistent and robust epithelial outgrowth, which was also abolished by trypsin/EDTA. Epithelial outgrowth generated by collagenase-isolated clusters was significantly larger in diameter and its single cells yielded more holoclones on 3T3 fibroblast feeder layers than that from dispase-isolated sheets. This new isolation method can be used for exploring how limbal epithelial stem cells are regulated by their native niche cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.