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Improved conditions for culture of biosynthetically active cockroach corpora allata

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Summary

Currently, short-term culture of insect corpora allata is most often performed in TC199. We now show that L-15B, a medium widely used in arthropod tissue culture, is superior to TC199 for both short- and long-term culture of cockroach corpora allata. In 3-h and 48-h incubations, juvenile hormone biosynthesis by corpora allata from Diploptera punctata was significantly higher in L-15B than in TC199. In addition, in both media, corpora allata activity was significantly improved by flotation of glands at the medium surface. Characteristics of L-15B responsible for its superiority were examined by comparison of gland activities in several TC199 formulations that had been modified in different ways to be more similar to L-15B. Adjusting the osmotic pressure of TC199 (288 mOsm/l) to near that of L-15B (362 mOsm/l) and D. punctata hemolymph (360 mOsm/l) significantly improved gland activity during the second 12 h of a 36-h incubation. Increasing the concentrations of amino acids, sugars, and organic acids in TC199 to the same levels as in L-15B significantly improved gland activity during both the second and third 12-h intervals of a 36-h incubation. These results suggest that L-15B is superior to TC199 because L-15B is isoosmotic with D. punctata hemolymph and because L-15B, like cockroach hemolymph, contains a high level of organic constituents. It is therefore more appropriate to use L-15B than TC199 for short-term in vitro assays of juvenile hormone biosynthesis and for extended corpora allata culture.

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Authors and Affiliations

  1. Department of Entomology, Box 7613, North Carolina State University, 27695-7613, Raleigh, North Carolina

    Glenn L. Holbrook & Coby Schal

  2. Institute of Life Sciences, National Tsing-Hua University, Hsinchu, Taiwan 30043, Republic of China

    Ann-Shyn Chiang

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  1. Glenn L. Holbrook
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  2. Ann-Shyn Chiang
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  3. Coby Schal
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Holbrook, G.L., Chiang, AS. & Schal, C. Improved conditions for culture of biosynthetically active cockroach corpora allata. In Vitro Cell.Dev.Biol.-Animal 33, 452–458 (1997). https://doi.org/10.1007/s11626-997-0063-9

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  • DOI: https://doi.org/10.1007/s11626-997-0063-9

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