Premium
The effect of several growth factors on keratocytes during the stromal wound repair in vitro
Author(s) -
GALLEGO P,
IBARESFRIAS L,
MARTINEZGARCIA C,
CANTALAPIEDRA R,
MERAYOLLOVES J
Publication year - 2010
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2010.205.x
Subject(s) - fetal bovine serum , stromal cell , platelet derived growth factor receptor , wound healing , cell growth , in vitro , growth factor , myofibroblast , microbiology and biotechnology , cell migration , fibroblast , andrology , platelet derived growth factor , transforming growth factor , cell culture , chemistry , biology , immunology , pathology , cancer research , medicine , biochemistry , fibrosis , receptor , genetics
Purpose To determine the effect of different growth factors during corneal stromal wound repair in vitro. Methods The effect of TGF‐β, FGFb and PDGF‐BB on proliferation, migration and differentiation was evaluated in human corneal keratocytes cultures. The same type of wound was performed in all cultures. After making wounds, keratocytes were cultured in different culture media: Dulbecco´s modified Eagle´s medium (DMEM/F12) or DMEM/F12 supplemented with TGF‐β, FGFb, PDGF‐BB or fetal bovine serum (FBS). Cultures were fixed after 4, 10 and 15 days, and then we studied the proliferation, differentiation and migration Results In serum‐free cultures, migration and cell proliferation rates were very low and the wounds closed after 15 days. These cells kept keratocyte morphology. The FBS addition produced fibroblast differentiation and the wounds were completely closed after 4 days. With this medium we observed the fastest rate of proliferation and cell migration of the study. TGF‐β treatment induced cell differentiation to myofibroblasts decreasing the rate of migration and cell proliferation so that the closure of the wound was very slow; 15 days after wounds performed these were not closed. Cell proliferation and migration were enhanced by FGFb and PDGF‐BB, for this reason the wounds were closed in 10 days. However both processes in PDGF‐BB supplemented cultures were more striking. Keratocytes morphology was maintained. Conclusion The effect of each growth factor signaling on keratocytes in vitro was involved in different events of wound repair, although the mixture of them in FBS improved the actions.