A method for the harvest, culture, and characterization of human adult atrial myocardial cells: correlation with age of donor.
Myocardial cell culture methods are now well established for animal and fetal human tissue. We present here a method for harvesting and culturing adult human atrial myocardiocytes. Cells are obtained from fresh atrial tissue normally discarded after being removed to cannulate the right atrium during open heart surgery. The atrial tissue is minced and then digested using collagenase. The single cell suspension is initially cultured in serum-containing growth medium, then transferred to defined medium, selective for myocardial cell growth. The cells are characterized by immunoperoxidase stains and transmission electron microscopy. The cultured cells stain positive for myoglobin, whereas control cultured fibroblasts and endothelial cells do not. Electron microscopy shows the presence of numerous myofibrils, Z-bodies, pleomorphic mitochondria, and secretory granules. The chronological age of the donor was an important factor in culturing the adult tissue, the younger tissue correlated with a higher success rate. This method provides a means for in vitro study of human adult myocardial cells and provides guidelines for appropriate atrial tissue to use.
Smith
D A
DA
William Beaumont Hospital, Royal Oak, Michigan 48073.
Glover
J L
JL
Townsend
L E
LE
Maupin
D E
DE
D000375:Aging; D002452:Cell Count; D002478:Cells, Cultured; D006325:Heart Atria / Q000166:cytology; D006801:Humans; D007124:Immunoenzyme Techniques; D008722:Methods; D008854:Microscopy, Electron; D009206:Myocardium / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D009211:Myoglobin / Q000032:analysis; D015388:Organelles / Q000648:ultrastructure; D014019:Tissue Donors
D009211:Myoglobin