Cyclic strain induces proliferation of cultured embryonic heart cells.
Embryonic heart cells undergo cyclic strain as the developing heart circulates blood to the embryo. Cyclic strain may have an important regulatory role in formation of the adult structure. This study examines the feasibility of a computerized cell-stretching device for applying strain to embryonic cardiocytes to allow measurement of the cellular response. A primary coculture of myocytes and a secondary culture of nonmyocytes from stage-31 (7 d) embryonic chick hearts were grown on collagen-coated membranes that were subsequently strained at 2 Hz to 20% maximal radial strain. After 24 h, total cell number increased by 37+/-6% in myocyte cocultures and by 26+/-6% in nonmyocyte cultures over unstrained controls. Lactate dehydrogenase and apoptosis assays showed no significant differences in cell viabilities between strained and unstrained cells. After 2 h strain, bromodeoxyuridine incorporation was 38+/-1.2% versus 19+/-0.2% (P < 0.01) in strained versus unstrained myocyte cocultures, and 35+/-2.1% versus 16+/-0.2% (P = 0.01) in nonmyocyte cultures. MF20 antibody labeling and periodic acid-Schiff (PAS) staining estimated the number of myocytes in strained wells as 50-67% larger than in control wells. Tyrosine phosphorylation may play a role in the cellular response to strain, as Western blot analysis showed an increase in tyrosine phosphorylation of two proteins with approximate molecular weights of 63 and 150 kDa within 2 min of strain. The results of this study indicate that embryonic chick cardiocytes can be cultured in an active mechanical environment without significant detachment and damage and that increased proliferation may be a primary response to strain.
Department of Pediatrics, University of Rochester School of Medicine and Dentistry, New York 14642, USA. ChristineE_Miller@urmc.rochester.edu
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Miller
K J
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Donlon
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Toia
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Wong
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Chess
D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D002642:Chick Embryo; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009206:Myocardium / Q000166:cytology* / Q000201:enzymology; D010766:Phosphorylation; D013314:Stress, Mechanical; D014443:Tyrosine / Q000378:metabolism
D014443:Tyrosine; D007770:L-Lactate Dehydrogenase; D001973:Bromodeoxyuridine