Isolation and Maintenance 



in Tissue Culture of 



Coelenterate Cell Lines 



John H. Phillips 



Department of Baeteriolofiij, University of California, Berkeley, California 



The in vitro cultivation of coelenterate tissues has been reported 

 before (1, 2). However, attempts at the maintenance of such cul- 

 tures for prolonged periods of time and serial transfer of cultured 

 material have not apparently met with success. In addition, the evi- 

 dence in support of true multiplication of cells has not been entirely 

 convincing. The methods that will be discussed have led to the 

 establishment of cell cultures from the anemone Anfhopleura ele- 

 gantissima. These cultures have been transferred twenty to thirty 

 times and have been under in vitro cultivation for more than a 

 year. In addition, eight of the cell lines have been through one 

 single cell cloning. The resulting clones of eight to thirty-two cells 

 have given rise to cultures containing 10*^ to 10" cells. 



First will be described the procedures which have been used 

 in isolation, cultivation and cloning of the cells, and this methodol- 

 ogy will be followed by a description of the cells and some of their 

 properties. 



A somewhat more detailed account of methods will soon be 

 published (5). All glassware and rubber stoppers were cleaned 

 by autoclaving in 0.1% NaoCO... (4). Glassware was wrapped in 

 aluminum foil and sterilized by dry heat. Rubber stoppers were 

 autoclaved in large Petri dishes. All nutrient solutions were steri- 

 hzed by filtration, using Millipore filters. The nutrient medium 

 that has been found to be most useful consists of 0.7% Edamine\ 



1 Sheffield Chemical Company, Inc., Norwich, N. Y. 



245 



