Microtextured substrata alter gene expression, protein localization and the shape of cardiac myocytes.
Many of the experiments designed to understand fundamental principles in cardiac physiology are performed in vitro using myocytes isolated from adult or neonatal hearts. However, these cells have probably lost some of their original properties in culture prior to study. Our objective is to recapitulate cardiac myocyte structure and function by growing cells on microtextured silicone substrata produced by photolithography and microfabrication techniques. Myocytes are plated on nontextured, micropegged (5 microm high), microgrooved (parallel grooves with a depth of 5 microm) or combination (micropegged and grooved) substrata. Myocytes plated on microtextured surfaces display a change in cell shape with an increase in myofibrillar height and a decrease in cell area. This shape change did not affect the stoichiometry of the myofibrillar proteins but did elicit microenvironmental remodeling of proteins that mechanically attach the cell to its surroundings. Cells terminate in a sarcomeric striation on the vertical interface of the peg whereas on nontextured surfaces they end in long nonstriated cables. Vinculin, a focal adhesion protein, was found to decrease in expression on combination surfaces as compared to nontextured substrata. A three-dimensional microtextured substratum appears to reintroduce a more physiological microarchitecture for tissue culture that may have potential uses in biological research as well as in tissue engineering and diagnostic applications.
Motlagh
Delara
D
Department of Physiology and Biophysics (M/C 901), University of Illinois at Chicago, 835 S. Wolcott Avenue, Chicago IL 60612-7342, USA.
Senyo
Sam E
SE
Desai
Tejal A
TA
Russell
Brenda
B
D000199:Actins / Q000378:metabolism*; D000818:Animals; D000831:Animals, Newborn; D002448:Cell Adhesion / Q000502:physiology; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D048429:Cell Size / Q000502:physiology; D002478:Cells, Cultured; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005786:Gene Expression Regulation / Q000502:physiology; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism*; D010781:Photography / Q000379:methods; D051381:Rats; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*; D016596:Vinculin / Q000378:metabolism*
D000199:Actins; D016596:Vinculin