Microfabricated grooves recapitulate neonatal myocyte connexin43 and N-cadherin expression and localization

Our objective is to alter the surface topography on which cardiac myocytes are grown in culture so that they more closely resemble their in vivo counterparts. Microtextured silicone substrata were made using photolithography and microfabrication techniques and then coated with laminin. Primary cardiac myocytes from newborn rats were plated on microgrooved and nontextured substrata. Myocytes were highly oriented on 5 microm grooves (69.8 +/- 2.0%) and significantly different, p < 0.0001, compared with randomly oriented cells grown on nontextured surfaces (2.9 +/- 0.95%; n = 19). Cells on shallower, 2 microm, grooves were slightly less well oriented (46.9 +/- 4.3%, n = 5, p < 0.001). The lateral spacings of the grooves were altered to examine changes in cell-to-cell contact by confocal immunocytochemistry and quantitative protein analysis. Connexin43 and N-cadherin were distributed around the perimeter of the myocytes plated on 10 x 5 x 5 microgrooved surfaces, similar to the localization found in the neonate. Connexin43 expression in cultures on 5 microm deep grooved substrata was equal to the neonatal heart, whereas it differed in nontextured surfaces. We conclude that it is necessary to combine groove depth (5 microm) and lateral ridge dimensions between grooves (5 microm) in order to recapitulate connexin43 and N-cadherin expression levels and subcellular localization to that of the neonate.