An explant culture system for the study of atrial development.
An explant culture system for the study of chick atrial development was tested using media of varying composition. Right atria from chick embryos of 13 days incubation were excised, placed on stainless steel mesh grids and cultured for 16-21 days in supplemented media. The results indicated that DMEM supplemented with 15% fetal bovine serum maintained the highest percentage (57%) of contractile atria in culture. Even atria cultured in serum free DMEM gradually adapted and became completely contractile over a 15 day period. Gel electrophoresis was performed on NP-40 detergent extracts of contracting explants cultured for 1, 4, 6, 8, and 10 days, and from 13, 15, 16, 18, 20 day embryos and 1 day old chicks. The percent of total protein of five separate protein bands in the range of 17,000 to 60,000 Daltons were plotted against the days in vivo/in vitro. There was a dramatic change in the protein profile during the first day of culture (an increase in concentration in the 60 kDa protein and decreases in the 17, 31, 37 and 50 kDa proteins). The in vivo atria showed a relatively large increase in the concentration of several of the proteins (17, 37, 50 kDA), profiled just prior to hatching with subsequent fall after hatching. A comparison between the biochemical data and the morphology of the explants indicate that although the explants display characteristics of contractility and typical cardiocyte structure, they were operative at a different functional level than their in vivo counterparts. The loss of homeostatic regulation and other in vivo embryonic factors could play a role in this observed change. This culture system provides an alternative to the dissociation method, and will be a useful model to study factors that influence atrial growth and development.
Call
T W
TW
Department of Anatomy, Medical College of Ohio, Toledo.
Lane
R D
RD
Arbogast
J
J
Morse
D E
DE
D000818:Animals; D002642:Chick Embryo; D003470:Culture Media / Q000494:pharmacology; D046508:Culture Techniques / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006325:Heart Atria / Q000187:drug effects / Q000196:embryology* / Q000648:ultrastructure; D013997:Time Factors
D003470:Culture Media