Abstract
Purpose: :
Retinal pigment epithelial (RPE) cell protein expression patterns differ in vivo and in vitro. In regular cell culture, RPE-65 is lost and the transdifferentiation marker smooth muscle actin is frequently expressed. Recent evidence suggests that substrate elasticity has a strong impact on several cell functions. Therefore, we assessed the effect of substrate elasticity on protein expression patterns in ARPE-19 cells.
Methods: :
ARPE-19 cells were plated on collagen-coated polyacrylamide gels of different elasticity or on regular glass or tissue culture plastic. Protein expression or transcription was assessed by Western Blot or qPCR, respectively. Staining for ki67 was used to examine the non-resting cell fraction.
Results: :
Highly elastic polyacrylamide substrates promote expression of the RPE-specific markers RPE-65 and Bestrophin-1 in vitro. The fraction of ARPE-19 cells expressing ki67 increases with substrate compliance.
Conclusions: :
ECM elasticity modulates protein expression in RPE cells. Elastic substrates promote the expression of RPE-specific markers. Thus, novel substrates of biomimetic elasticity may significantly enhance RPE cell propagation in vitro.
Keywords: extracellular matrix • retinal pigment epithelium