Embryonic development of an endoparasitoid,Microplitis croceipes (Hymenoptera: Braconidae) in cell line-conditioned media |
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Authors: | Stephen M Ferkovich Herbert Oberlander Charles Dillard Eddie Leach |
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Institution: | (1) Insect Attractants, Behavior, and Basic Biology Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, P.O. Box 14565, 32604 Gainesville, Florida |
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Abstract: | Summary Embryos of the parasitoidMicropolitis croceipes develop from pregerm band stage to first larval instar in cell culture medium conditioned by a cell line (IPLB-LdFB) derived
from fat body from an atypical hostLymantria dispar. However, the percentage of eggs that develop normally to the first larval instar stage is significantly less than for those
maintained in IPL-52B medium conditioned with host fat body tissue. Therefore, we examined the capacity of five insect cell
lines to promote growth and development of pregerm band eggs in five media, IPL-52B, TC-199, TC-100, Grace’s, and ExCell 400.
The developmental response ofM. croceipes was dependent both on the cell line and the cell culture medium used. TC-100, TC-199, and Grace’s media promoted development
to the germ band stage without the need for conditioning with host tissue. IPL-52B supported development to the germ hand
stage when a defined lipid concentrate was added. In IPL-52B medium, the IPLB-LdFB cell line promoted a significantly higher
number of eggs developing to germ band relative to the other cell lines; however, none of the cell line-conditioned IPL-52B
medium significantly stimulated egg hatch relative to the control medium. None of the cell line-conditioned Grace’s media
had a significant effect on eggs attaining germ band stage compared with the Grace’s control medium. However, Grace’s medium
conditioned with the IAL-TND1 and IPLB-LdFB cell lines promoted development beyond germ band, resulting in a significantly
higher percentage of hatching eggs than the Grace’s control medium. Although the BCIRL-HZ-AMI cell line, which is derived
from the parasitoid’s typical host, did not induce hatch in either IPL-52B medium or Grace’s medium, it promoted hatch in
TC-199 and Excell 400 media. Fat body taken from the same species that the cell lines were derived from was a better predictor
of a cell line’s embryotrophic activity in Grace’s medium rather than in IPL-52B medium. Thus, the composition of the medium
and the species and tissue type of the cell line source must be evaluated interactively to determine optimal conditions for
promoting development ofM. croceipes in vitro. |
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Keywords: | cell line media germ band larval instar fat body hatch lipid |
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