A simple device to apply equibiaxial strain to cells cultured on flexible membranes.
The biomechanical environment to which cells are exposed is important to their normal growth, development, interaction, and function. Accordingly, there has been much interest in studying the role of biomechanical forces in cell biology and pathophysiology. This has led to the introduction and even commercialization of many experimental devices. Many of the early devices were limited by the heterogeneity of deformation of cells cultivated in different locations of the culture plate membranes and were also attached with complicated technical/electronic efforts resulting in a restriction of the reproducibility of these devices. The objective of this study was to design and build a simple device to allow the application of dose-dependent homogeneous equibiaxial static stretch to cells cultured on flexible silicone membranes to investigate biological and biomedical questions. In addition, cultured neonatal rat atrial cardiomyocytes were stretched with the proposed device with different strain gradients. For the first time with this study we could demonstrate that stretch up to 21% caused dose-dependent changes in biological markers such as the calcineurin activity, modulatory calcineurin-interacting protein-1, voltage-gated potassium channel isoform 4.2, and voltage-gated K(+) channel-interacting proteins-2 gene expression and transient outward potassium current densities but not the protein-to-DNA ratio and atrial natriuretic peptide mRNA. With both markers mentioned last, dose-dependent stretch alterations could only be achieved with stretch up to 13%. The simple and low-cost device presented here might be applied to a wide range of experimental settings in different fields of research.
Rana
Obaida R
OR
Univ. Hospital RWTH Aachen, Dept. I of Internal Medicine, Division of Cardiology, Pulmonary and Vascular Diseases, Pauwelsstrasse 30, Aachen, Germany. orana@ukaachen.de
Zobel
Carsten
C
Saygili
Esra
E
Brixius
Klara
K
Gramley
Felix
F
Schimpf
Thomas
T
Mischke
Karl
K
Frechen
Dirk
D
Knackstedt
Christian
C
Schwinger
Robert H G
RH
Schauerte
Patrick
P
Saygili
Erol
E
D000818:Animals; D000831:Animals, Newborn; D009320:Atrial Natriuretic Factor / Q000235:genetics / Q000378:metabolism; D019703:Calcineurin / Q000378:metabolism; D018929:Cell Culture Techniques / Q000295:instrumentation*; D048429:Cell Size; D002478:Cells, Cultured; D004867:Equipment Design; D006325:Heart Atria / Q000378:metabolism; D006984:Hypertrophy; D047908:Intracellular Signaling Peptides and Proteins; D051605:Kv Channel-Interacting Proteins / Q000235:genetics / Q000378:metabolism; D008422:Materials Testing; D008564:Membrane Potentials; D008567:Membranes, Artificial; D032383:Myocytes, Cardiac / Q000378:metabolism* / Q000473:pathology; D018583:Pliability; D011188:Potassium / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D051672:Shal Potassium Channels / Q000235:genetics / Q000378:metabolism; D012828:Silicones / Q000737:chemistry*; D013314:Stress, Mechanical; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D047908:Intracellular Signaling Peptides and Proteins; C496758:Kcnip2 protein, rat; D051605:Kv Channel-Interacting Proteins; D008567:Membranes, Artificial; C548050:RCAN1 protein, rat; D012333:RNA, Messenger; D051672:Shal Potassium Channels; D012828:Silicones; D014157:Transcription Factors; D009320:Atrial Natriuretic Factor; D019703:Calcineurin; D011188:Potassium