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Long-term primary culture of secretory cells of Bothrops jararaca venom gland for venom production in vitro

Nature Protocols volume 1, pages 2763–2766 (2006)Cite this article

Abstract

This protocol details the optimal conditions to establish a long-term primary culture of secretory cells from the venom gland of the Bothrops jararaca snake. Furthermore, these conditions allow the production and secretion of venom into the culture medium. Snake venom is a rich source of active molecules and has been used for bioprospection studies. However, obtaining enough venom from snakes is a major obstacle. Secretory cells of venom glands are capable of producing active toxins. Therefore, a culture of secretory cells is a good in vitro system to acquire the venom of snakes without capturing the animal from the wild. The protocol described here provides a rapid (∼4 h) and reproducible means of producing sufficient amounts of snake venom for biological investigations.

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Figure 1: Dissection of venom gland from Bothrops jararaca snake.
Figure 2: Secretory cells of snake venom gland in culture.
Figure 3: Venom produced in culture causes hemorrhagic lesions.

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Acknowledgements

The authors thank Dr. D.O. Quissell for suggestions on setting up the methodology, Stella Furlan for technical assistance in animal procedures, Thiago Macedo de Abreu Hortêncio for photographing the dissection of venom glands and Dr. Colin Jamora for a careful review of this manuscript. This work was supported by CNPq (478700-2004-0) and FAPESP (02/00422-8, 06/54188-7).

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Authors and Affiliations

  1. Laboratório de Farmacologia, Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, 05503-900, Brazil

    Norma Yamanouye & Celine Marie Kerchove

  2. Laboratório de Imunopatologia, Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, 05503-900, Brazil

    Ana Maria Moura-da-Silva

  3. Laboratório de Biologia Celular, Instituto Butantan, Avenida Vital Brazil, 1500, São Paulo, 05503-900, Brazil

    Sylvia M Carneiro

  4. Departamento de Fisiologia, Instituto de Biociências, Universidade de São Paulo, Rua do Matão, travessa 14, São Paulo, 05508-900, Brazil

    Regina P Markus

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  1. Norma Yamanouye
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Correspondence to Norma Yamanouye.

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Yamanouye, N., Kerchove, C., Moura-da-Silva, A. et al. Long-term primary culture of secretory cells of Bothrops jararaca venom gland for venom production in vitro. Nat Protoc 1, 2763–2766 (2006). https://doi.org/10.1038/nprot.2006.423

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