TY - JOUR
T1 - Micro-perfusion for cardiac tissue engineering
T2 - Development of a bench-top system for the culture of primary cardiac cells
AU - Khait, Luda
AU - Hecker, Louise
AU - Radnoti, Desmond
AU - Birla, Ravi K.
N1 - Funding Information:
We would like to thank the Section of Cardiac Surgery for financial support toward the development and testing of the perfusion system. RB and LK were supported by funds from the Section of Cardiac Surgery. Financial support for LH was provided by the American Heart Association (pre-doctoral fellowship grant # 0615557Z). We would like to thank Radnoti Glass Technology Inc for providing us with several prototypes for various components of the perfusion system.
PY - 2008/5
Y1 - 2008/5
N2 - Tissue-engineered constructs have high metabolic requirements during in vitro culture necessitating the development of micro-perfusion systems to maintain high functional performance. In this study, we describe the design, fabrication, and testing of a novel micro-perfusion system to support the culture of primary cardiac cells. Our system consists of a micro-incubator with independent stages for 35-mm tissue culture plates with inflow/outflow manifolds for fluid delivery and aspiration. A peristaltic pump is utilized for fluid delivery and vacuum for fluid aspiration. Oxygen saturation, pH, and temperature are regulated for the media while temperature is regulated within the micro-incubator, fluid reservoir, and oxygenation chamber. Validation of the perfusion system was carried out using primary cardiac myocytes, isolated from 2- to 3-day-old neonatal rat hearts, plated on collagen-coated tissue culture plates. Two million cells/plate were used and the perfusion system was run for 1 h (without the need for a cell culture incubator) while controls were maintained in a standard cell culture incubator. We evaluated the cell viability, cell adhesion, total protein, total RNA, and changes in the expression of SERCA2 and phospholamban using RT-PCR, with N = 6 for each group. We found that there was no significant change in any variable during the 1-h run in the perfusion system. These studies served to demonstrate the compatibility of the perfusion system to support short-term culture of primary cardiac cells.
AB - Tissue-engineered constructs have high metabolic requirements during in vitro culture necessitating the development of micro-perfusion systems to maintain high functional performance. In this study, we describe the design, fabrication, and testing of a novel micro-perfusion system to support the culture of primary cardiac cells. Our system consists of a micro-incubator with independent stages for 35-mm tissue culture plates with inflow/outflow manifolds for fluid delivery and aspiration. A peristaltic pump is utilized for fluid delivery and vacuum for fluid aspiration. Oxygen saturation, pH, and temperature are regulated for the media while temperature is regulated within the micro-incubator, fluid reservoir, and oxygenation chamber. Validation of the perfusion system was carried out using primary cardiac myocytes, isolated from 2- to 3-day-old neonatal rat hearts, plated on collagen-coated tissue culture plates. Two million cells/plate were used and the perfusion system was run for 1 h (without the need for a cell culture incubator) while controls were maintained in a standard cell culture incubator. We evaluated the cell viability, cell adhesion, total protein, total RNA, and changes in the expression of SERCA2 and phospholamban using RT-PCR, with N = 6 for each group. We found that there was no significant change in any variable during the 1-h run in the perfusion system. These studies served to demonstrate the compatibility of the perfusion system to support short-term culture of primary cardiac cells.
KW - 3D constructs
KW - Cardiac cells
KW - Heart muscle
KW - Perfusion
KW - RT-PCR
KW - Tissue engineering
UR - http://www.scopus.com/inward/record.url?scp=43249103373&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=43249103373&partnerID=8YFLogxK
U2 - 10.1007/s10439-008-9459-2
DO - 10.1007/s10439-008-9459-2
M3 - Article
C2 - 18274906
AN - SCOPUS:43249103373
SN - 0090-6964
VL - 36
SP - 713
EP - 725
JO - Annals of Biomedical Engineering
JF - Annals of Biomedical Engineering
IS - 5
ER -