Evaluation of Heat Shock Protein 90 and Heat Shock Constitutive Protein7o Expression in Cultured Limbal Stem Cells During Air Exposure (Pages: 0-0)


Mohamadi P *, Ebrahimi M , Baharvand H ,

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Objective: Heat shock proteins (Hsps) are highly conserved proteins which have important roles in protection against stress, signaling pathway and differentiation, embryonic development and chaperon activity. The aim of this study is investigation of HSP90 and HSC70 expression before and after air exposure in cultivated limbal stem cells on denuded amniotic membrane(AM) and plastic dish(PD). Materials and Methods: Limbal biopsies were taken from normal human limbus (Central Eye Bank of Iran), and were cultured as an explant on amniotic membrane and plastic dish. Cells were exposed to the air after 14 days of culture. The expression of proposed limbal stem cells markers (p63,ABCG2),corneal markers(K3/12,Connexin43) and also HSPs(90,hsc70) were analyzed by RT-PCR, immunocytochemistry and flowcytometry pre and post air exposure at day 14 (before air exposure) and 30(after air exposure) respectively. Results: ABCG2 positive cells with high expression of p63 and low expression of K3/K12 and Connexin43 were considered as limbal stem cells while ABCG2 negative cells with low expression of p63 and high expression of K3/K12,Connexin43 were assumed as corneal cells. Our data showed that HSPs expressed strongly at mRNA level before and after air exposure while they expressed in different pattern at protein level. It was interested that HSP90 proteins were expressed weakly in limbal stem cells at day 14 on both groups, but its expression increased after air exposure at day 30(p