Characterization and comparison of human limbal explant cultures grown under defined and xeno-free conditions
Language English Country England, Great Britain Media print-electronic
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
29928900
DOI
10.1016/j.exer.2018.06.019
PII: S0014-4835(18)30184-2
Knihovny.cz E-resources
- Keywords
- Explant cultures, Fibrin, Human platelet lysate, Human serum, Limbal epithelial stem cells, Limbal stem cell deficiency,
- MeSH
- Coated Materials, Biocompatible MeSH
- Biomarkers metabolism MeSH
- Cell Differentiation MeSH
- Cell Culture Techniques MeSH
- Tissue Donors MeSH
- Adult MeSH
- Fibrin pharmacology MeSH
- Culture Media MeSH
- Cells, Cultured MeSH
- Real-Time Polymerase Chain Reaction MeSH
- Middle Aged MeSH
- Humans MeSH
- Limbus Corneae cytology metabolism MeSH
- Cell Count MeSH
- Feeder Cells MeSH
- Cell Proliferation physiology MeSH
- Epithelium, Corneal cytology metabolism MeSH
- Aged MeSH
- Serum * MeSH
- Blood Platelets * MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Aged MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
- Names of Substances
- Coated Materials, Biocompatible MeSH
- Biomarkers MeSH
- Fibrin MeSH
- Culture Media MeSH
Human limbal epithelial cells (LECs) intended for treatment of limbal stem cell deficiency are commonly cultivated on a 3T3 feeder layer with complex culture medium supplemented with fetal bovine serum (FBS). However, FBS is a xenogeneic component containing poorly characterised constituents and exhibits quantitative and qualitative lot-to-lot variations. Human limbal explants were plated on untreated or fibrin coated plastic plates and cultured in two non-xenogeneic media (supplemented with either human serum or platelet lysate only). Our aim was to find out whether the characteristics of harvested LEC cultures are comparable to those of LEC cultivated in the gold standard - FBS-supplemented complex medium. The growth kinetics, cell proliferation, differentiation, stemness maintenance, apoptosis and contamination by other cell types were evaluated and compared among these conditions. In all of them LECs were successfully cultivated. Stemness was preserved in both xeno-free media. However, cells cultured with human serum on the fibrin-coated plates had the highest growth rate and cell proliferation and very low fibroblast-like cell contamination. These data suggest that xeno-free cell culture conditions can replace the traditional FBS-supplemented medium and thereby provide a safer protocol for ex vivo cultured limbal stem cell transplants.
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