Characterization and comparison of human limbal explant cultures grown under defined and xeno-free conditions
Jazyk angličtina Země Anglie, Velká Británie Médium print-electronic
Typ dokumentu srovnávací studie, časopisecké články, práce podpořená grantem
PubMed
29928900
DOI
10.1016/j.exer.2018.06.019
PII: S0014-4835(18)30184-2
Knihovny.cz E-zdroje
- Klíčová slova
- Explant cultures, Fibrin, Human platelet lysate, Human serum, Limbal epithelial stem cells, Limbal stem cell deficiency,
- MeSH
- biokompatibilní potahované materiály MeSH
- biologické markery metabolismus MeSH
- buněčná diferenciace MeSH
- buněčné kultury MeSH
- dárci tkání MeSH
- dospělí MeSH
- fibrin farmakologie MeSH
- kultivační média MeSH
- kultivované buňky MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé středního věku MeSH
- lidé MeSH
- limbus corneae cytologie metabolismus MeSH
- počet buněk MeSH
- podkladové buňky MeSH
- proliferace buněk fyziologie MeSH
- rohovkový epitel cytologie metabolismus MeSH
- senioři MeSH
- sérum * MeSH
- trombocyty * MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- biokompatibilní potahované materiály MeSH
- biologické markery MeSH
- fibrin MeSH
- kultivační média 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.
Citace poskytuje Crossref.org
