Abstract 1284: Laser Patterned Adult Cardiomyocyte Cell-Culture Model

Communication between neighboring cells is crucial for the understanding of the physiology and pathology of tissues. However, due to the complexity of in vivo environments, it is hard to investigate this communication directly on tissues isolated from the body. Therefore, it is important to construct a cell-culture model that will replicate the most relevant characteristics of that tissue. Native cardiomyocytes have an aligned arrangement in which neighboring cardiomyocytes are electrically and mechanical coupled through contact junctions. When adult cardiomyocytes are placed into a culture dish, the cells will be randomly oriented and die quicker due to the lack of proper aligned cell-cell connections. To address this issue, we have implemented our laser cell micropatterning system to create an adult cardiomyocyte culturing model with aligned rows of cells connected end to end. In this abstract, we describe the experimental procedure to achieve the laser alignment of adult cardiomyocytes and report the preliminary observation on the constructed cell-culture model. To harvest adult cardiomyocytes from rats, we placed the rats under general anesthesia using intraperitoneal injection of Sodium Pentobarbital (2%, containing 1:200 Heparin 45mg/kg). The chest cavity was then opened and the anterior vena cava and postcaval vein were quickly cut. We then perfused Krebs Ringer’s buffer (KRB) solution with high potassium (KCl) into the ascending aorta and the cardiac cells were collected by the followed coronary perfusion with collagenase. Our laser cell alignment system is based on a weakly focused laser beam that exerts a radial force trapping a cell into the beam and an axial force that propels the cell in the direction of the beam’s propagation. Individual cardiomyocytes were injected through a microinjection system into the cell deposition chamber. The laser beam controlled by a computer system, turned the cell to a vertical orientation and moved to the appropriate position during deposition. Following cells were placed one by one to form a contact aligned array. Our data have shown that the laser cell micropatterning system can effectively align adult myocytes onto a 3D collagen substrate to construct an aligned adult cardiomyocyte cell-culture model.