Abstract
As part of a bioassay approach to investigate ovarian development and function, primary cell cultures were derived from Penaeus monodon ovaries at various stages of maturation. These cultures were established in modified Grace's or modified 2× L-15 media. Various supplements including growth factors, vitamins and minerals were trailed. Four morphologically different types of cells (epithelioid, fibroblastic, rounded, and epithelioid with large nuclei) were maintained for up to 17 months. Epithelioid cells grew best in modified Grace's medium but were generally short-lived (less than two months). Fibroblast-like cells formed confluent monolayers in modified 2× L-15 medium, were passaged three times and survived for 17 months. In other cultures, millions of rounded cells migrated from tissue. They survived for prolonged periods (up to ten months), either loosely attached to the flask or suspended in the medium. A change in dominant cell type from fibroblastic to epithelioid was observed in some cultures after three or nine months incubation. These epithelioid cells which had very large nuclei, grew to confluence but could not be sub-cultured. It is noteworthy that the rounded cells and the epithelioid cells with the large nuclei both produced vitellogenin in protein-free media.
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References
Bell TA, Lightner DV (1988). A Handbook of Normal Penaeid Shrimp Histology, pp 36–37. Baton Rouge, LA: World Aquaculture Society.
Chang ES, Brody MD (1989). Crustacean organ and cell culture. In: Maramorosch K, Sato GH (ed), Advances in Cell Culture, vol 7, pp 19–86. New York: Acad Press.
Chen S-N, Chi SC, Kou GH, Liao IC (1986). Cell culture from tissues of grass prawn, Penaeus monodon. Fish Pathol 21: 161–166.
Chen S-N, Jong KJ, Kou GH (1988). Cell cultures from haematopoietic tissue and ovary of penaeid shrimp, Penaeus monodon. In: Kuroda Y, Kurstak E, Maramorosch K (eds), Invertebrate and Fish Tissue Culture: Proceedings of the Seventh International Conference on Invertebrates and Fish Tissue Culture, Japan, 1987, pp 195–198. Tokyo: Springer-Verlag.
Chen S-N, Jong KJ, Kou GH (1989). Cell cultures derived from tissues of penaeid shrimp, Penaeus penicillatus, and hard clam, Meretrix lusoria. In: Mitsuhashi J (ed), Invertebrate Cell System Applications, vol II, pp 253–262. Boca Raton, Florida: CRC Press.
Ellender RD, Najafabadi AK, Middlebrooks BL (1992). Observations on the primary culture of haemocytes of Penaeus. J Crustacean Biol 12: 178–185.
Hsu Y-L, Yang Y-H, Chen Y-C, Tung M-C, Wu J-L, Engelking MH, Leong JC (1995). Development of an in vitro subculture system for the oka organ (Lymphoid tissue) of Penaeus monodon. Aquaculture 136: 43–55.
Huberman A, Aguilar MB, Quackenbush LS (1995). A neuropeptide family from sinus gland of the Mexican crayfish, Procambarus bouvieri (Ortmann). Aquaculture 135: 149–160.
Ke H, Liping W, Yumei D, Shuji Z (1990). Studies on a cell culture from the hepatopancreas of the oriental shrimp, Penaeus orientalis Kishinouye. Asian Fisheries Sci 3: 299–307.
Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Lee C-Y, Watson RD (1995). In vitro study of vitellogenesis in the blue crab (Callinectes sapidus): Site and control of vitellin synthesis. J Exp Zool 271: 364–372.
Luedeman RA, Lightner DV (1992). Development of an in vitro primary cell system from the penaeid shrimp, Penaeus stylirostris and Penaeus vannamei. Aquaculture 101: 205–211.
Machii A, Wada KT, Awaji M, Nakamura HK, Townsley SJ (1988). Some characteristics of cells of the midgut gland and chytrids from primary cultures of the Penaeus japonicus. In: Kuroda Y, Kurstak E, Maramorosch K (eds), Invertebrate and Fish Tissue Culture: Proceedings of the Seventh International Conference on Invertebrates and Fish Tissue Culture, Japan, 1987, pp 11–14. Tokyo: Springer-Verlag.
Munderloh UG, Kurtti TJ (1989). Formulation of medium for tick cell culture. Exp Appl Acarol 7: 219–229.
Najafabadi AK, Ellender RD, Middlebrooks BL (1992). Analysis of shrimp haemolymph and ionic modification of a penaeus cell culture formulation. J Aquat Animal Hlth 4: 143–148.
Quackenbush LS (1989). Vitellogenesis in the shrimp, Penaeus vannamei: in vitro studies of the isolated hepatopancreas and ovary. Comp Biochem Physiol 94B: 253–261.
Quackenbush LS (1992). Yolk synthesis in the marine shrimp, Penaeus vannamei. Comp Biochem Physiol 103A: 711–714.
Quackenbush LS, Keeley LL (1988). Regulation of vitellogenesis in the fiddler crab, Uca pugilator. Biol Bull 175: 321–331.
Tapay LM, Lu Y, Brock JA, Nadala Jnr EC, Loh PC (1995). Transformation of primary cultures of shrimp (Penaeus stylirostris) lymphoid (Oka) organ with simian virus-40 (T) antigen. Proc Soc Exp Biol Med 209: 73–78.
Tong S-L, Miao H-Z (1996). Attempts to initiate cell cultures from Penaeus chinensis tissues. Aquaculture 146: 151–157.
Toullec J-Y, Chikhi M, van Wormhoudt A (1992). In vitro protein synthesis and α amylase activity in F cells from the hepatopancreas of Palaemon serratus (Crustacea; Decapoda). Experientia 48: 272–277.
Toullec J-Y, Crozat Y, Patrois J, Porcheron P (1996). Development of primary cell cultures from the penaeid shrimps Penaeus vannamei and P. indicus. J Crustacean Biol 16(4): 643–649.
Toullec J-Y, Dauphin-Villemant C (1994). Dissociated cell suspensions of Carinus maenas Y-organs as a tool to study ecdysteroid production and its regulation. Experientia 50: 153–158.
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Fraser, C.A., Hall, M.R. Studies on primary cell cultures derived from ovarian tissue of Penaeus monodon. Methods Cell Sci 21, 213–218 (1999). https://doi.org/10.1023/A:1009851809288
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DOI: https://doi.org/10.1023/A:1009851809288