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Characterization and comparison of human limbal explant cultures grown under defined and xeno-free conditions.
Brejchova, Kristyna; Trosan, Peter; Studeny, Pavel; Skalicka, Pavlina; Utheim, Tor Paaske; Bednar, Jan; Jirsova, Katerina.
Afiliação
  • Brejchova K; Research Unit for Rare Diseases, Clinic of Paediatrics and Adolescent Medicine, 1st Faculty of Medicine, Charles University, Ke Karlovu 2, 128 08 Prague 2, Czech Republic; Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, 1st Faculty of Medicine, Charles
  • Trosan P; Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, Albertov 4, 128 00 Prague 2, Czech Republic.
  • Studeny P; Ophthalmology Department of 3rd Medical Faculty and University Hospital Kralovske Vinohrady, Srobárova 1150/50, 100 34 Prague 10, Czech Republic.
  • Skalicka P; Research Unit for Rare Diseases, Clinic of Paediatrics and Adolescent Medicine, 1st Faculty of Medicine, Charles University, Ke Karlovu 2, 128 08 Prague 2, Czech Republic; Department of Ophthalmology, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, U nemocnice
  • Utheim TP; Department of Medical Biochemistry, Oslo University Hospital, Kirkeveien 166, 0407 Oslo, Norway; Department of Plastic and Reconstructive Surgery, Oslo University Hospital, Kirkeveien 166, 0407 Oslo, Norway.
  • Bednar J; Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, Albertov 4, 128 00 Prague 2, Czech Republic.
  • Jirsova K; Laboratory of the Biology and Pathology of the Eye, Institute of Biology and Medical Genetics, 1st Faculty of Medicine, Charles University and General University Hospital in Prague, Albertov 4, 128 00 Prague 2, Czech Republic; Ophthalmology Department of 3rd Medical Faculty and University Hospital K
Exp Eye Res ; 176: 20-28, 2018 11.
Article em En | MEDLINE | ID: mdl-29928900
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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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Guideline / Qualitative_research Limite: Adult / Aged / Humans / Middle aged Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Guideline / Qualitative_research Limite: Adult / Aged / Humans / Middle aged Idioma: En Ano de publicação: 2018 Tipo de documento: Article