| Title |
Cultivation and Characterization of Cornea Limbal Epithelial Stem Cells on Lens Capsule in Animal Material-Free Medium
|
|---|---|
| Published in |
PLOS ONE, October 2012
|
| DOI | 10.1371/journal.pone.0047187 |
| Pubmed ID | |
| Authors |
Réka Albert, Zoltán Veréb, Krisztián Csomós, Morten C. Moe, Erik O. Johnsen, Ole Kristoffer Olstad, Bjørn Nicolaissen, Éva Rajnavölgyi, László Fésüs, András Berta, Goran Petrovski |
| Abstract |
A simple, reproducible, animal-material free method for cultivating and characterizing cornea limbal epithelial stem cells (LESCs) on human lens capsule (LC) was developed for future clinical transplantation. The limbal tissue explants (2 × 2 × 0.25 mm) were harvested from 77 cadavers and expanded ex vivo on either cell culture plates or LC in medium containing human serum as the only growth supplement. Cell outgrowth at the edge of the explants was observed within 24 hours of cultivation and achieved viable outgrowth (>97% viability as measured by MTT assay and flow cytometry) within two weeks. The outgrowing cells were examined by genome-wide microarray including markers of stemness (p63α, ABCG2, CK19, Vimentin and Integrin α9), proliferation (Ki-67), limbal epithelial cells (CK 8/18 and 14) and differentiated cornea epithelial cells (CK 3 and 12). Immunostaining revealed the non-hematopoietic, -endothelial and -mesenchymal stem cell phenotype of the LESCs and the localization of specific markers in situ. Cell adhesion molecules, integrins and lectin-based surface carbohydrate profiling showed a specific pattern on these cells, while colony-formation assay confirmed their clonal potency. The LESCs expressed a specific surface marker fingerprint (CD117/c-kit, CXCR4, CD144/VE-Cadherin, CD146/MCAM, CD166/ALCAM, and surface carbohydrates: WGA, ConA, RCA, PNA and AIL) which can be used for better localization of the limbal stem cell niche. In summary, we report a novel method combining the use of a medium with human serum as the only growth supplement with LC for cultivating, characterizing and expanding cornea LESCs from cadavers or alternatively from autologous donors for possible treatment of LESC deficiency. |
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 66 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United Kingdom | 1 | 2% |
| Turkey | 1 | 2% |
| Colombia | 1 | 2% |
| Unknown | 63 | 95% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 13 | 20% |
| Researcher | 12 | 18% |
| Student > Master | 9 | 14% |
| Student > Postgraduate | 6 | 9% |
| Other | 5 | 8% |
| Other | 12 | 18% |
| Unknown | 9 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 20 | 30% |
| Agricultural and Biological Sciences | 16 | 24% |
| Biochemistry, Genetics and Molecular Biology | 9 | 14% |
| Engineering | 2 | 3% |
| Chemical Engineering | 1 | 2% |
| Other | 2 | 3% |
| Unknown | 16 | 24% |