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Establishment of a lepidopteran hybrid cell line by use of a biochemical blocking method

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We report here on the establishment of aCydia pomonella (Cp) hybrid cell line IZD-Cp 4/13. As there have been no reports on somatic cell fusion involving lepidopteran cell lines so far, we had to develop an appropriate fusion procedure. We first tried—but without much success—to obtain HAT (hypoxanthine, aminopterin and thymidine)- or TAM (thymidine, adenine, and methotrexate)-selectable strains of the threeCydia pomonella cell lines IZD-Cp 2202, IZD-Cp 0508 and Cp 169. We then tried and succeeded in developing a fusion procedure based on the use of biochemically blocked permanent cells as one partner in the fusion. Biochemically inhibited IZD-Cp 2202 cells and embryonic Cp cells were hybridized by polyethyleneglycol treatment. The cells of the hybrid cell line IZD-Cp 4/13 differ from the permanent parental cells (cell line IZD-Cp 2202) with respect to morphology, DNA content isoenzyme patterns, and response to challenge with theChoristoneura murinana nuclear polyhedrosis virus.

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References

  1. Chen, T. R. In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 333258 stain. Exp. Cell Res. 104:255–262; 1977.

    Article  PubMed  CAS  Google Scholar 

  2. Darai, G.; Zoeller, L.; Fluegel, R. M.; et al. Mink lung cells as a tool for detection of Mycoplasma hyorhinis contamination in cell cultures and virus stocks. In Vitro 19:7–15; 1983.

    Article  PubMed  CAS  Google Scholar 

  3. Ellender, R.; Middlebrooks, B. L.; Toom, P. M. Simple isoelectric focusing method to quantitate LDH isoenzyme in cultured cells for cell characterization. TCA Manual 3(2):593–596; 1977.

    Article  Google Scholar 

  4. Goldsby, R. A.; Osborne, B. A.; Simpson, E.; et al. Hybrid cell lines with T-cell characteristics. Nature 267:707–708; 1977.

    Article  PubMed  CAS  Google Scholar 

  5. Hink, W. F.; Ellis, B. J. 3. Establishment and characterization of two new cell lines (Cp-1268 and Cp-169) from the codling moth,Carpocapsa pomonella. Curr. Top. Microbiol. Immunol. 55:19–28; 1971.

    PubMed  CAS  Google Scholar 

  6. Hunter, L. Glucose-6-phosphate dehydrogenase isoenzymes in cultured human cell lines: Separation by isoelectric focusing. Anal. Biochem. 101:78–87; 1980.

    Article  PubMed  CAS  Google Scholar 

  7. Kobayashi, Y.; Asada, M.; Higuchi, M.; et al. Human T cell hybridomas producing lymphokines. I. Establishment and characterization of human T cell hybridomas producing lymphotoxin and migration inhibitory factor. J. Immunol. 128:2714–2718; 1982.

    PubMed  CAS  Google Scholar 

  8. Kontiainen, S.; Simpson, E.; Bohrer, E.; et al. T-cell lines producing antigen-specific suppressor factor. Nature 274:477–480; 1978.

    Article  PubMed  CAS  Google Scholar 

  9. Littlefield, J. W. Selection of hybrids from matings of fibroblasts in vitro and their presumed recombinants. Science 145:709–710; 1964.

    Article  PubMed  CAS  Google Scholar 

  10. Maskos, C. B.; Miltenburger, H. G. Isoenzym-Analyse von Insektenzellen durch isoelektrische Fokussierung in Duennschicht-Polyacrylamid-Gelen. Mitt. Dtsch. Ges. Allg. Angew. Ent. 4:44–47; 1983.

    Google Scholar 

  11. Miltenburger, H. G.; David, P.; Mahr, U.; et al. Ueber die Erstellung von Lepidopteren-Dauerzellinien und die in vitro-Replikation von insektenpathogenen Biren. I. Mamestra brassicae L. (Kohleule)-Zellinien und NPV-Replikation. Z. Ang. Entomol. 82:306–323; 1977.

    Google Scholar 

  12. Miltenburger H. G.; Naser, W. L.; Harvey, J. P.; et al. The cellular substrate: A very important requirement for baculovirus in vitro replication. Naturforsch 39c:993–1002; 1984.

    CAS  Google Scholar 

  13. Saga, S.; Tsutsui, J.; Imai, M.; et al. The enhanced expression of cellular fibronectin in mouse-mouse somatic cell hybrids. Exp. Cell Res. 141:478–483; 1982.

    Article  PubMed  CAS  Google Scholar 

  14. Simms, N. Methods for detection of enzymatic activity after electrophoresis on polyacrylamide gel in Drosophila species. Nature 207:757–758; 1965.

    Article  Google Scholar 

  15. Wyss, C. TAM selection of Drosophila somatic cell hybrids. Somatic Cell Genet. 5:29–37; 1979.

    Article  PubMed  CAS  Google Scholar 

  16. Wyss, C. Loss of ecdysterone sensitivity of a Drosophila cell line after hybridization with embryonic cells. Exp. Cell Res. 125:121–126; 1979.

    Article  Google Scholar 

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Authors and Affiliations

  1. Institute of Zoology, Technical University Darmstadt, Schnittspahnstr. 3, D-6100, Darmstadt, Federal Republic of Germany, FRG

    H. G. Miltenburger, W. L. Naser & M. G. Schliermann

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  1. H. G. Miltenburger
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  2. W. L. Naser
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  3. M. G. Schliermann
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This work was supported by the Bundesministerium fuer Forschung und Technologie, Bonn, FRG.

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Miltenburger, H.G., Naser, W.L. & Schliermann, M.G. Establishment of a lepidopteran hybrid cell line by use of a biochemical blocking method. In Vitro Cell Dev Biol 21, 433–438 (1985). https://doi.org/10.1007/BF02620830

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  • DOI: https://doi.org/10.1007/BF02620830

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