CULTURE OF CELLS AND TISSUES DERIVED FROM XENOPUS LAEVIS 619 



N° 31. H. Moser, L Hadji-Azimi and S. Slatkine. - Culture of 

 cells and tissues derived from the south African frog Xenopus 

 laevis (Daudin). 1 ' 2 (8 figures and 4 tables.) 



Station de Zoologie expérimentale, Université de Genève, Suisse. 



Abstract 



The procédures for layer-cultures of Xenopus laevis cells, tissues and organ 

 fragments adapted by or developed in our laboratory are described, and some 

 preliminary observations with explanted Xenopus cells are reported. 



Introduction 



In order to study with in vitro methods certain problems of Amphibian 

 development, vve established, during the past year, a tissue culture laboratory 

 at the Station de Zoologie expérimentale (University of Geneva). With the aid 

 of modem methods (Moser, H., 1960) we initiated a project to produce primary 

 cell cultures derived from Xenopus laevis (Daudin) and to establish eventually 

 permanent cell strains thereof. 



Xenopus cells and small tissue fragments isolated and explanted according 

 to the procédures currently in use with mammalian and avian cells generally 

 respond well to in vitro conditions if the tonicity of the nutrient média and wash 

 solutions and the incubation température are sufficiently lowered, and if the 

 concentration of cells in the inocula for the initiation of primary cultures is 

 increased. An exception are the very large and fragile cells of early embryonic 

 stages which do not stand well the standard enzymatic methods of cell dispersion 

 (Rafferty, K. A., in press). According to our expérience an osmolarity cor- 

 responding to 80% of that one of mammalian liquid média 3 and an incubation 



1 Supported by grant No. 4411 from the Fond National de la Recherche Scientifique 

 Suisse, and by a grant from the A. and J. CLARAZ Foundation. 



2 The authors wish to express their gratitude to Professor Michail Fischberg for his contin- 

 ued personal interest in this work and for the generous support he has given for if s exécution. 

 The authors also wish to thank for the technical help rendered by Mlle. Lotte Voll, Mlle. Mireille 

 Maye, Mlle. Dr. Janina Kapinska and Mlle. Walburg Riemann. 



3 Balls and Ruben (1966) report the successful growth of primary Xenopus laevis cell 

 cultures in a médium composed of 50% Leibovitz-L15, 40% H 2 and 10% fetal bovine sérum, 

 that is, in a growth médium of an osmolarity of only 60% of that of mammalian cell culture 

 média. Our expérience has clearly shown that this médium is hypotonie for the majority of cells 

 derived from Xenopus laevis tissues and causes rapid lysis of large numbers of cells in primary 

 explant cultures. However, undoubtedly certain (selected) Anuran cell types survive in growth 

 média of 60% osmolarity, as this is demonstrated by the fact that J. J. Freed has been able to 

 establish, in this type of growth médium, an autodiploid cloned cell strain of Rana pipiens 

 (ICR-RPH-134cl). 



Rev. Suisse de Zool., T. 75, 1968 



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