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Protocol no. 106
EMBRYONIC MYOCARDIAL MYOCYTE REAGGREGATION CULTURES AS A MODEL FOR CARDIOTOXICITY

Reaggregates of isolated chick embryo myocardial myocytes are used in a simple tissue culture based test as a model for the cardiotoxicity of compounds.

CONTACT

Dr. C.D.N. Toseland, Dr. L. K. Earl
Smith Kline Beecham Pharmaceuticals
Unilever
The Frythe Environmental Safety Laboratory
Welwyn Colworth House Herts.
AL6 9AR Sharnbrook UK
Bedford,
MK44 1LQ, UK
Tel: +44 (0)1438 782670
Tel: +44 (0)1234 222013
Fax: +44 (0)1438 782582
Fax: +44 (0)1234 222122

RATIONALE

Direct-acting cardiotoxic agents may be distinguished from agents causing cardiotoxicity by indirect mechanisms by their more marked effects in chick myocardial myocyte reaggregate cultures.

BASIC PROCEDURE

Myocardial myocytes are isolated from 10 day old chick embryo hearts and cultured in 24-well plates for 3-4 before exposure to the test substance. Inhibition of spontaneous beating activity, leakage of lactate dehydrogenase, and changes in cell morphology can be used as toxic endpoints.

TEST STATUS

No formal international validation has taken place, but results from two different laboratories have been consistent.

CHEMICALS TESTED

Allylamine HCl Diallylamine Digoxin Doxorubicin Hydralazine Isoprenaline n-Methyl allylamine Pinacidil

REFERENCES

  1. Boor P.J., and Ferrans V.J. (1982) Ultrastructural alterations in allylamine-induced cardiomyopathy. Early lesions. Laboratory Investigations 47: 76-86.
  2. Clusin W.T. (1981) The mechanical activity of chick embryonic myocardial cell aggregates. J. Physiol. 320: 149-179.
  3. Earl L.K., Kesingland K., Davis K.P., Brocklehurst S.R., and Jones H.B. (1992) Allylamine toxicity in embryonic myocardial myocyte reaggregate cultures: The role of extracellular metabolism by benzylamine oxidase. Toxic. in Vitro 6: 405-416.
  4. Kesingland K., Earl L.K., Roberts J.C., and Jones H.B. (1991) Allylamine toxicity in embryonic myocardial myocyte reaggregate cultures. Toxic. in Vitro 5: 145-156.
  5. Seaman C.W., Toseland C.D.N., Maile P.A., Francis I., and White D.J. (1994) A study of the cardiotoxic potential of pharmaceutical compounds on chick myocardial myocyte reaggregate cultures. Toxic. in Vitro 8: 543-544.
  6. Shirhatti V., George M., Chenery R, and Krishna G. (1986) Structural requirements for inducing cardiotoxicity by anthracycline antibiotics: studies with neonatal rat cardiac myocytes in culture. Toxicol. Appl. Pharmacol. 84: 173-191.
  7. Sperelakis N., and Bkaily G. (1987) Cultured cell models for studying problems in cardiac toxicology. in In Vitro Methods in Toxicology (eds. C. Atterwill and C.E. Steele). Cambridge University Press; Cambridge, pp. 59-108.
  8. Toraason M., Luken M.E., Breitenstein M., Krueger J.A., and Biagini R.E. (1989) Comparative toxicity of allylamine and acrolein in cultured myocytes and fibroblasts from neonatal rat heart. Toxicology 56: 107-117.

IP-106 November 94