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Long-Term Cultures of Human Cornea Limbal Explants Form 3D Structures Ex Vivo - Implications for Tissue Engineering and Clinical Applications
- Source :
- PLoS ONE, Vol 10, Iss 11, p e0143053 (2015), PLoS ONE
- Publication Year :
- 2015
- Publisher :
- Public Library of Science (PLoS), 2015.
-
Abstract
- Long-term cultures of cornea limbal epithelial stem cells (LESCs) were developed and characterized for future tissue engineering and clinical applications. The limbal tissue explants were cultivated and expanded for more than 3 months in medium containing serum as the only growth supplement and without use of scaffolds. Viable 3D cell outgrowth from the explants was observed within 4 weeks of cultivation. The outgrowing cells were examined by immunofluorescent staining for putative markers of stemness (ABCG2, CK15, CK19 and Vimentin), proliferation (p63α, Ki-67), limbal basal epithelial cells (CK8/18) and differentiated cornea epithelial cells (CK3 and CK12). Morphological and immunostaining analyses revealed that long-term culturing can form stratified 3D tissue layers with a clear extracellular matrix deposition and organization (collagen I, IV and V). The LESCs showed robust expression of p63α, ABCG2, and their surface marker fingerprint (CD117/c-kit, CXCR4, CD146/MCAM, CD166/ALCAM) changed over time compared to short-term LESC cultures. Overall, we provide a model for generating stem cell-rich, long-standing 3D cultures from LESCs which can be used for further research purposes and clinical transplantation.
- Subjects :
- Pathology
medicine.medical_specialty
Primary Cell Culture
Gene Expression
lcsh:Medicine
Biology
Limbus Corneae
Extracellular matrix
Tissue engineering
Antigens, CD
Cornea
medicine
ATP Binding Cassette Transporter, Subfamily G, Member 2
Humans
Vimentin
lcsh:Science
ALCAM
Cell Proliferation
Multidisciplinary
Tissue Engineering
Stem Cells
Tumor Suppressor Proteins
lcsh:R
Epithelium, Corneal
Cell Differentiation
Epithelial Cells
Epithelium
Culture Media
Neoplasm Proteins
Transplantation
medicine.anatomical_structure
Ki-67 Antigen
Cell culture
Keratins
ATP-Binding Cassette Transporters
lcsh:Q
Autopsy
Collagen
sense organs
Stem cell
Biomarkers
Research Article
Transcription Factors
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 10
- Issue :
- 11
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....e868506f4e6db802c44ee2298a14a59a