Factors promoting the establishment of primary cultures of liver cells from Xenopus larvae Article Received: 21 March 1977 Accepted: 19 May 1977 DOI:
10.1007/BF00848385 Cite this article as: Wahli, W. & Weber, R. Wilhelm Roux' Archiv (1977) 182: 347. doi:10.1007/BF00848385 Summary
The requirements for establishment and survival of primary cultures of larval amphibian liver cells were investigated.
Plating efficiency was found to be enhanced by a collagen substrate, by diluted conditioned medium from an adult
Xenopus kidney cell line and by high initial cell densities. Plating efficiency was highest at a tonicity of 165–220 mOsm/kg. In cultures with undiluted conditioned medium the increase in cell number was 50–60% greater than in controls, where it was about 2-fold between day 3 and 6 of culture. Conditioned medium from the Xenopus kidney cell line is assumed to contain at least two components, which are effective at different concentrations and stimulate either plating efficiency and cell aggregation or cell proliferation.
In cultures without collagen sheets, cell flattening is greatly reduced, indicating that cell shape is also dependent upon the substrate.
Key words Xenopus larvae Liver Primary culture Plating efficiency Cell proliferation References
Arthur, E., Balls, M.: Amphibian cells in vitro. I. Growth of Xenopus cells in a soft agar medium and on agar surface. Exp. Cell Res.
, 113–118 (1971)
Balinsky, J.B., Goetzer, T.L., Mattheyre, F.J.: The effect of thyroxin and hypertonic environment on the enzymes of the urea cycle in Xenopus laevis. Comp. Biochem. Physiol.
, 83–95 (1972)
Balls, M., Worley, R.S.: Amphibian cells in vitro. II. Effect of variations in medium osmolarity on a permanent cell line isolated from Xenopus. Exp. Cell Res.
, 333–336 (1973)
Balsamo, J., Lilien, J.: Embryonic cell aggregation: Kinetics and specificity of binding of enhancing factors. Proc. nat. Acad. Sci. (Wash.)
, 727–731 (1974)
Bennet, R.P., Kriegstein, H., Glenn, I.S.: Thyroxine stimulation of ornithine transcarbamylase activity and protein synthesis in tadpole (Rana catesbeiana) liver in organ culture. Biochem. Biophys. Res. Commun.
, 412–417 (1969)
Blatt, L.M., Kim, K.-H., Cohen, P.P.: The effect of thyroxine on ribonucleic acid synthesis by premetamorphic tadpole liver cell suspensions. J. Biol. Chem.
, 4801–4807 (1969)
Calderon, J., Unanue, E.R.: Two biological activites regulating cell proliferation found in cultures of peritoneal exudate cells. Nature
, 359–361 (1975)
Chen, P.S.: Patterns and metamorphic changes of serum proteins in Amphibia. Wilhelm Roux's Archiv
, 132–149 (1970)
Dulak, N.C., Temin, H.M.: A partially purified polypeptide fraction from rat liver cell conditioned medium with multiplication-stimulating activity for embryo fibroblasts. J. Cell. Physiol.
, 153–160 (1973)
Garber, B., Moscona, A.A.: Reconstruction of brain tissue from cell suspensions. II. Specific enhancement of aggregation of embryonic cerebral cells by supernatant from homologous cultures. Develop. Biol.
, 235–243 (1972)
Gasche, P.: Zur Frage des Angriffspunktes des Thiouracil. Experientia
, 24–26 (1946)
Godsell, P.: The cell cycle of Xenopus laevis cells in monolayer culture. Exp. Cell Res.
, 433–436 (1974)
Hauschka, S.D.: Clonal aspects of muscle development and the stability of the differentiated state. In: “The stability of the differentiated state” (H. Ursprung, ed.), pp. 37–57, Berlin: Springer-Verlag 1968
Kistler, A., Weber, R.: Enzyme patterns in mitochondria of eggs, liver, and skeletal muscle during larval development of
. Develop. Biol.
, 236–247 (1974)
Koch, K., Leffert, H.L.: Growth control of differentiated fetal rat hepatocytes in primary monolayer culture. VI. Studies with conditioned medium and its functional interactions with serum factors. J. Cell Biol.
, 780–791 (1974)
Leford, B.E., Frieden, E.: Albumin synthesis during induced and spontaneous metamorphosis in the bullfrog Rana catesbeiana. Develop. Biol.
, 187–197 (1973)
Leffert, H.L., Paul, D.: Studies on primary cultures of differentiated foetal liver cells. J. Cell Biol.
, 559–568 (1972)
McGuire, E.J.: Intercellular adhesive selectivity. II. Properties of embryonic chick liver cell-cell adhesion. J. Cell Biol.
, 90–100 (1976)
Michalopoulos, G., Pitot, H.C.: Primary culture of parenchymal liver cells on collagen membranes. Morphological and biochemical observations. Exp. Cell Res.
, 70–78 (1975)
Monnickendam, A.M., Balls, M.: Amphibian organ culture. Experientia
, 1–17 (1973)
Moscona, A.A., Moscona, M.H.: Inhibition of cell aggregation in vitro by puromycin. Exp. Cell Res.
, 697–702 (1966)
Nagano, H., Shimada, T., Shukuya, R.: Increase in serum albumin during bullfrog metamorphosis. J. Biochem.
, 1307–1309 (1973)
Nieuwkoop, P.D., Faber, J.: Normal table of Xenopus laevis (Daudin). Amsterdam: North Holland Publishing Co. 1956
Paul, J.: Cell and tissue culture, fourth edition, pp. 73–74. London: Churchill Livingstone 1973
Shaffer, B.M.: The isolated Xenopus laevis tail: A preparation for studying the central nervous system and metamorphosis in culture. J. Embryol. exp. Morph
Shambaugh, G.E., Balinsky, J.B., Cohen, P.P.: Synthesis of carbamoylphosphate synthetase in Amphibian liver in vitro. J. Biol. Chem.
, 5295–5308 (1969)
Solursh, M., Reiter, R.S.: Long-term cell culture of two differentiated cell types from the liver of larval and adult Xenopus. Z. Zellforsch.
, 457–469 (1972)
Takahashi, K., Okada, T.S.: An analysis of the effect of conditioned medium upon the cell culture at low density. Develop. Growth Differ.
, 65–77 (1970)
Takahashi, K., Okada, T.S.: Separation of two factors affecting the aggregation kinetics from the conditioned medium. Develop. Growth Differ.
, 15–24 (1971)
Tata, J.R.: Requirement for RNA and protein synthesis for induced regression of the tadpole tail in organ culture. Develop. Biol.
, 77–94 (1966)
Watanabe, K.: Changes in the capacity for clonal growth and differentiation in vitro of the vertebral cartilage cells with embryonic development. II. Vitalizing effect of conditioned medium on the cells of younger embryos. Develop. Growth Differ.
, 107–118 (1971)
Weber, R.: Behaviour and properties of acid hydrolases in regressing tails of tadpoles during spontaneous and induced metamorphosis in vitro. In: “Lysosomes”. Ciba Foundation Symposium (A.V.S. de Reuck and M.P. Cameron, eds.), pp. 282–300. London: Churchill Ltd 1963
Wood, G.C., Keech, M.K.: Collagen precipitation rate. Biochem. J.
, 588 (1968)