Skip to main content
Log in

In vitro differentiation of rabbit blastocyst cells

  • Regular Papers
  • Published:
In Vitro Cellular & Developmental Biology Aims and scope Submit manuscript

Summary

Six- and seven-day post-coitus (p.c.) rabbit embryos have been cultured in an attempt to establish a trophectodermal cell line. Results indicate that cells with epithelial characteristics (i.e. positive staining for cytokeratin) will survive in culture until Passage 3. At that time a fibroblastlike cell becomes predominant. In addition, we have found that the presence of the inner cell mass is required for embryo explants often results in the development of cells that spontaneously contract. These cells stain positively for myosin, which indicates that they may be precardiac cells. Maximum diastolic potential was −59±1.2 mV and the threshold potential was −53±2.3 mV. Spontaneously contracting cells did not respond to atropine, acetylcholine, epinephrine, isoproterenol, or propranolol. Action potential seems to be a result of an inward calcium current, because the beating rate is decreased in a dose-related manner with the calcium channel blocker verapamil, whereas the voltage-sensitive sodium channel blocker tetrodotoxin was without effect.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Babinet, C.; Bordenave, G. R. Chimaeric rabbits from immunosurgically-prepared inner-cell-mass transplantation. J. Embryol. Exp. Morphol. 60:429–440; 1980.

    PubMed  CAS  Google Scholar 

  • Baer, M.; Best, P. M.; Reuter, H. Voltage-dependent action of tetrodotoxin in mammalian cardiac muscle. Nature 263:344–345; 1976.

    Article  PubMed  CAS  Google Scholar 

  • Beier, H. M. Oviductal and uterine fluids. J. Reprod. Fertil. 37:221–237; 1974.

    Article  PubMed  CAS  Google Scholar 

  • Bell, J. E.; Begg, K. E.; Sin, Y., et al. Neutral amino acid influx in developing rabbit blastocysts. Am. J. Physiol. 251:C285-C292; 1986.

    PubMed  CAS  Google Scholar 

  • Benos, D. J. Ouabain binding to preimplantation rabbit blastocysts. Dev. Biol. 83:69–78; 1981a.

    Article  PubMed  CAS  Google Scholar 

  • Benos, D. J. Development changes in epithelial transport characteristics of preimplantation rabbit blastocysts. J. Physiol. 316:191–202; 1981b.

    PubMed  CAS  Google Scholar 

  • Benos, D. J.; Biggers, J. D. Sodium and chloride co-transport by preimplantation rabbit blastocysts. J. Physiol. 342:23–33; 1983.

    PubMed  CAS  Google Scholar 

  • Cole, R. J.; Edwards, R. G.; Paul, J. Cytodifferentiation in cell colonies and cell strains derived from cleaving ova and blastocysts of the rabbit. Exp. Cell. Res. 37:501–504; 1965.

    Article  PubMed  CAS  Google Scholar 

  • Freshney, R. I. Culture of animal cells—A manual of basic technique. New York: Alan R. Liss; 1983.

    Google Scholar 

  • Friedman, W. F.; Pool, P. E.; Jacobowitz, D., et al. Sympathetic innervation of the developing rabbit heart. Circ. Res. 23:25–32; 1968.

    PubMed  CAS  Google Scholar 

  • Gardner, R. L. An investigation of inner cell mass and trophoblastic tissue following their isolation from the mouse blastocyst. J. Embryol. Exp. Morphol. 28:277–312; 1972.

    Google Scholar 

  • Ilgren, E. B. The control of trophoblastic growth in the guinea pig. J. Embryol. Exp. Morphol. 60:405–418; 1980.

    PubMed  CAS  Google Scholar 

  • Rizzino, A. Early mouse embryos produce and release factors with transforming growth factor activity. In vitro 21:531–536; 1985.

    CAS  Google Scholar 

  • Rizzino, A. Defining the roles of growth factors during early mammalian development. In: Bavister, B. D., ed. The mammalian preimplantation embryo. Regulation of growth and differentiation in vitro. New York: Plenum Press; 1987:151–174.

    Google Scholar 

  • Robinson, D. H.; Kirk, K. L.; Benos, D. J. Uteroglobin transport in rabbit blastocyst. FASEB. J. 2:A308; 1988.

    Google Scholar 

  • Rowlands, I. W.; Weir, B. J. Mammals: non-primate eutherians. In: Lamming, G. E., ed. Marshall's physiology of reproduction, vol 1. New York: Churchill Livingstone; 1984:455–658.

    Google Scholar 

  • Spies, H. G.; Chappel, S. C. Mammals: non-human primates. In: Lamming, G. E., ed. Marshall's physiology of reproduction, vol. I. New York: Churchill Livingstone; 1984:659–712.

    Google Scholar 

  • Spitzer, N. C. Ion channels in developments. Ann. Rev. Neurosci. 2:363–397; 1979.

    Article  PubMed  CAS  Google Scholar 

  • Summers, P. M.; Taylor, C. T.; Hearn, J. P. Characteristics of trophoblastic tissue derived from in vitro culture of preimplantation embyros of the common marmoset monkey. Placenta 8:411–422; 1987.

    PubMed  CAS  Google Scholar 

  • Van Mierop, L. H. S. Location of the pacemaker in chick embrvo heart at the time of initiation of heartbeat. Am. J. Physiol. 212:407–415; 1967.

    PubMed  Google Scholar 

  • Whyte, A.; Ockleford, C. D.; Wooding, F. D. P., et al. Characteristics of cells derived from the girdle region of the preimplantation blastocyst of the donkey. Cell Tissue Res. 246:343–352; 1986.

    Article  PubMed  Google Scholar 

  • Woods, W. T.; Urthaler, F.; James, T. N. Spontaneous action potentials of cells in the canine sinus node. Circ. Res. 39:76–82; 1976.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This work was supported by grants HD21302, HD07069, DK31091, and HL37320 from the National Institutes of Health, Bethesda, MD, with additional support from a University of Alabama at Birmingham Cardviovascular Research and Training Center Award.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gargus, E.B., Robinson, D.H., Bubien, J.K. et al. In vitro differentiation of rabbit blastocyst cells. In Vitro Cell Dev Biol 25, 435–441 (1989). https://doi.org/10.1007/BF02624628

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02624628

Key words

Navigation