ArticleInvestigative ophthalmology & visual science2020
A Comparison of Methods for Isolation of Limbal Niche Cells: Maintenance of Limbal Epithelial Stem/Progenitor Cells.
Article in Investigative ophthalmology & visual science, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 14 citations in OpenAlex.
- 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
- Assessment of the toxic effect of benzalkonium chloride on human limbal stem cells.Scientific reports · 2025Article
- Transdifferentiation of rat keratinocyte progenitors to corneal epithelial cells by limbal niche via the STAT3/PI3K/AKT signaling pathway.Stem cell research & therapy · 2025Article
- Posterior Limbal Mesenchymal Stromal Cells Promote Proliferation and Stemness of Transition Zone Cells: A Novel Insight Into Corneal Endothelial Rejuvenation.Investigative ophthalmology & visual science · 2025Article
- Isolation andBio-protocol · 2022Article
- Limbal Niche Cells and Three-Dimensional Matrigel-Induced Dedifferentiation of Mature Corneal Epithelial Cells.Investigative ophthalmology & visual science · 2022Article
- Efficient Isolation and Functional Characterization of Niche Cells from Human Corneal Limbus.International journal of molecular sciences · 2022Article
- Progress of EGFL6 in angiogenesis and tumor development.International journal of clinical and experimental pathology · 2022Review
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Purpose: Limbal niche cells (LNCs) play a vital role in the maintenance of limbal epithelial stem/progenitor cells (LESCs). Four methods have been reported to isolate and expand LNCs: digestion by collagenase alone (C-LNC), collagenase following dispase removal of the limbal epithelium (DC-LNC), dissection of dispase-isolated limbal epithelial sheets (D-LNC), and explant cultures of limbal stromal tissues (Ex-LNC). This study aimed to isolate LNCs using those four methods and to compare their capacity to maintain LESCs. Methods: LNCs were isolated from the rat corneal limbus by the following methods: C-LNC, DC-LNC, D-LNC, and Ex-LNC. Quantitative real-time PCR and immunofluorescence staining were used to analyze the expression of embryonic stem cell (ESC) markers. The ability to maintain LESCs was assessed on the basis of colony-forming capacity and the expression of progenitor, proliferation, and differentiation markers in three-dimensional (3D) Matrigel and Transwell systems. Notch signaling of LESCs supported by different LNCs in Transwell inserts was analyzed by quantitative real-time PCR. Results: DC-LNCs exhibited lower expression of CK12 during isolation and expansion. Among P4-expanded LNCs, DC-LNCs expressed significantly higher levels of Sox2, Oct4, Nanog, and N-cadherin than C-LNCs, D-LNCs, and Ex-LNCs. Compared with other LNCs, DC-LNCs were more effective in maintaining LESCs with higher holoclone-forming efficiency, greater expression of ΔNp63α and Ki67, and lower expression of CK12. DC-LNCs were also more capable of downregulating Notch signaling of LESCs. Conclusions: DC-LNCs were more effective in expressing ESC markers and maintaining LESCs compared to other LNCs. This study identifies an optimal method for the isolation of LNCs in tissue engineering and ocular surface reconstruction.
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