Abstract
This paper reports for the first time, the Primary cell culture of hepatopancreas from edible crab Scylla serrata using crab saline, L-15 (Leibovitz), 1 × L-15 + crab saline, 2 × L-15 + crab saline, 3 × L-15 and citrate buffer without any serum. We could isolate and maintain E (Embryonalzellen), F (Fibrenzellen), B (Blasenzellen), R (Restzellen) and G (Granular cells). Upon seeding the hepatopancreatic E, F, B, and R cells showed different survival pattern over time than granular cells. A modified L-15 (3×) medium supported the best survival of hepatopancreatic E, F B, and R cells in in-vitro culture. However granular cells could be maintained for 184 days with L-15 (1×) + crab saline. Fetal bovine serum was not effective additive and hampered cell viability in present study.
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Al-Mohanna SY, Nott JA (1985) Mitotic E-and secretory F-cells in the hepatopancreas of the shrimp Penaeus Semisulcatus (Crustacea: Decapoda). J mar Biol ASS UK 65:901–910
Al-Mohanna SY, Nott JA (1987) R-cells and the digestive cycle in Penaeus semisulcatus (Crustacean: Decapoda). Mar Biol (Berl) 95:129–137. doi:10.1007/BF00447494
Ballard TA, Roer RD, Dillaman RM (1993) Long-term culture of integumental explants from premolt blue crabs, Callinectes sapidus. J Tissue Cult Methods 15:11–14. doi:10.1007/BF02387283
Chen S-N, Chi S-C, Kou G-H, Liao IC (1986) Cell cultures from tissues of Grass Prawn, Penaeus monodon. Fish Pathol 21:161–166
Chen S-N, Shih H-H, Kou G-H (1995) Primary cell cultures from tissues of penaeid shrimps and their susceptibilities to monodon-type baculovirus (MBV). Rep Fish Dis Res 16:1–14
Chen SN, Wen CM (1999) Establishment of cell lines derived from oyster, Crassostrea gigas Thunberg and hard clam, Meretrix lusoria Roding. Methods Cell Sci 21:183–192. doi:10.1023/A:1009829807954
Frerichs GN (1996) In vitro culture of embryonic cells from the freshwater prawn Macrobrachium rosenbergii. Aquaculture 143:227–232. doi:10.1016/0044-8486(96)01281-1
Hsu Y-L, Yang Y-H, Chen Y-C, Tung M-C, Wu J-L, Engelking MH et al (1995) Development of an in vitro subculture system for prawn tissues. Asian Fish Soc Spec Pub 10:161–170
Icely JD, Nott JA (1992) Digestion and absorption: digestive system and associated organs. In: Harrison FW, Humes AG (eds) Microscopic anatomy of invertebrates, vol 10. Decapod crustacea, Wiley-Liss, New York, pp 147–201
Johnston DJ, Alexander CG, Yellowhees D (1998) Epithelial cytology and function in the digestive gland of Thenus orientalis (Decapoda, scyllaridae). J Crust Biol 18(12):271–278. doi:10.2307/1549320
Ke H, Liping W, Yumei D, Shuji Z (1990) Studies on cell culture from the hepato-pancreas of the oriental shrimp, Penaeus orientalis Kishinouye. Asian Fish Sci 3:299–307
Klinbunga S, Boonyapakdee A, Pratoomchat B (2000) Genetic diversity and species-diagnostic markers of mud crabs (Genus Scylla) in Eastern Thailand determined by RAPD analysis. Mar Biotechnol 2:180–187
Luedeman RA (1990) Development of an in vitro primary cell cultures from the penaeid shrimp Penaeus stylirostris and Penaeus vannamei and evaluation of a potential application. Masters Thesis. The University of Arizona, Tucson, AZ
Mulford AL, Austin B (1998) Development of primary cell cultures from Nephrops norvegicus. Methods Cell Sci 19:269–275. doi:10.1023/A:1009787223797
Mulford AL, Lyng F, Mothersill C, Austin B (2001) Development and characterization of primary cell cultures from the hematopoietic tissues of the Dublin Bay prawn, Nephrops norvegicus. Methods Cell Sci 22:265–275. doi:10.1023/A:1017971618398
Nadala EC, Lu Y, Loh PC (1993) Primary culture of lymphoid, nerve and ovary cells from Penaeus stylirostris and Penaeus vannamei. In Vitro Cell Dev Biol Anim 29A:620–622. doi:10.1007/BF02634546
Leigh Owens, Jan Smith (1999) Early attempts at production of prawn cell lines. Methods Cell Sci 21:207–211. doi:10.1023/A:1009806606562
Peponnet F, Quiol JM (1971) Cell cultures of crustacea Arachnida and merostomacea. In: Vago C (ed) Invertebrate tissue culture. vol I pp 341–359
Rosenthal J, Diamant A (1990) In vitro primary cell culture from penaeus semisulcatus. In: perkins FO, Cheng TC (eds) Pathology in Marine Science. Academic Press, San Diego, pp 7–13
Sousa LG, Elena I, Cuartas , Petriella AM (2005) Fine structural analysis of the epithelial cells in the hepatopancreas of Palaemonetes argentinus (Crustacea, Decapoda, Caridea) in intermoult. Biocell 29(1):25–31
Tong S-L, Miao H-Z (1996) Attempts to initiate cell cultures from Penaeus chinensis tissues. Aquaculture 147:151–157. doi:10.1016/S0044-8486(96)01386-5
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 Crust Biol 16:643–649. doi:10.2307/1549183
Uma A, Prabhakar TG, Koteeswaran A, Ravikumar G (2002) Establishment of primary cell culture from hepatopancreas of Penaeus monodon for the study of whitespot syndrome virus (WSSV). Asian Fish Sci 15:365–370
Zilli L, Schiavone R, Scordella G, Zonno V, Verri T, Storelli C et al (2003) Changes in cell type composition and enzymatic activities in the hepatopancreas of Marsupenaeus japonicus during the moulting cycle. J Comp Physiol [B] 173:355–363. doi:10.1007/s00360-003-0348-6
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The authors acknowledge the receipts of financial assistances from Department of Biotechnology, New Delhi under the project number BT/PR8029/AAQ/03/287/2006.
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Sashikumar, A., Desai, P.V. Development of primary cell culture from Scylla serrata . Cytotechnology 56, 161–169 (2008). https://doi.org/10.1007/s10616-008-9152-1
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DOI: https://doi.org/10.1007/s10616-008-9152-1