Perfusionists in the world of tissue engineering--a quick and easy method for safe isolation of huge quantities of porcine cardiomyocytes.
Isolation of huge quantities of primary cells from whole organs like the heart becomes increasingly important, especially for the emerging research field of tissue engineering and regenerative medicine. This study deals with the isolation of pig cardiomyocytes, in contrast to the standard mouse or rat models, because we aimed to draw attention to the species, which are genetically more closely related to the human organism. The bigger operative and veterinary expenditure of the pig-heart model can only be justified by a technique that supplies a big amount of qualitative high-grade cardiomyocytes. In our model, the quality is guaranteed by protection of the heart, already in situ, by a cardioplegia and a careful application of collagenase to soften the tissue. The construction of a new apparatus which includes enormous costs was not necessary, since the perfusion equipment was realized from two commercially available HLMsets, which were modified and connected to each other. Our model makes it possible to rinse the whole myocardium, which leads to a better output than models that only prepare a part of the myocardium around a coronary artery. The careful harvesting of high-grade cardiomyocytes is an important source of successful cell cultures to be used for numerous experimental applications, may reduce animal experiments and, additionally, represents a chance for perfusionists to become an important partner in interdisciplinary research projects.
Neumann
Bernd
B
Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital, Tuebingen, Germany.
Guo
Ketai
K
Paul
Angela
A
Greiner
Tim O
TO
Graessle
Angelika
A
Walter
Christian
C
Walker
Tobias
T
Scheule
Albertus
A
Ziemer
Gerhard
G
Wendel
Hans P
HP
D000758:Anesthesia; D000818:Animals; D001011:Aorta; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases; D004867:Equipment Design; D006324:Heart Arrest, Induced; D006355:Heart-Lung Machine; D032383:Myocytes, Cardiac / Q000166:cytology*; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods*; D013045:Species Specificity; D034421:Sus scrofa; D023822:Tissue Engineering / Q000379:methods*
D017364:Collagenases