Worm Breeder's Gazette 2(2): 2

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Techniques for the Large-scale Axenic Culture and Aging of Turbatrix aceti and Possible Applications to C. elegans

W.F. Hieb, M. Rothstein

For several years we have been studying age-related changes in 
enzymes isolated from the free-living nematode Turbatrix aceti.  This 
has required the large-scale cultivation, synchronization and aging of 
nematodes in axenic, chemically defined media.
Organisms are grown in one liter Roux bottles containing 50 ml of 
medium for 14 to 16 days ant then harvested by filtration with a 
fritted glass Buechner funnel.  Small individuals are separated by 
'screening' through stainless steel filter cloth and transferred to a 
medium containing fluorodeoxyuridine (FUDR) plus uridine (100  g/ml 
and 150  g/ml, respectively).  FUDR inhibits reproduction, but not the 
life-span of the nematodes and thereby permits the development and 
ageing of cultures without contamination by newborn organisms.  Growth 
of the organisms in length is nearly normal during the aging process.  
Turbatrix can also be synchronized by increasing the incubation 
temperature to 36 C or by repeated removal of small larvae from 
cultures.  Biochemical characteristics of nematodes obtained by these 
methods appear to be similar to those of FUDR-treated worms.
The yield of organisms in mixed culture on a wet weight basis is 
approximately 2 grams per 50 ml of medium in a period of two weeks.  
Our maximum incubator capacity is 500 Roux bottles, resulting in a 
potential production of a kilogram of nematodes per fortnight.  
Quantities available from aged worms are only 10 to 20% of this figure 
due to losses in screening and to the limitations of the worm 
concentration which is permissible in 'ageing' cultures.
Based on our experience with Turbatrix and Caenorhabditis briggsae 
and various methods in the published literature, it is reasonable to 
conclude that C.  elegans can be synchronized and aged under similar 
conditions.  An advantage to using this organism is its ability to 
grow in cultures which are aerated by mechanical agitation.  The use 
of rotary shaker cultures permits a greater yield of organisms and 
more economy of space than static cultures.  Unfortunately, T.  aceti 
will not grow under the former conditions.
The culture of nematodes in completely defined medium offers an 
advantage for certain biological studies.  However, C.  elegans has a 
requirement for an unidentified growth factor present in soy-peptone.  
Elucidation of the structure of this material is therefore of 
importance if we are to fully utilize this organism for ageing studies.
We also believe that the cultivation of C.  elegans in an axenic 
system has definite potential for developmental biology and genetics.