caa a22 4500 445292504 CHVBK 20180317142532.0 cr unu---uuuuu 170323e20101001xx s 000 0 eng 10.1007/s11626-010-9343-x doi (NATIONALLICENCE)springer-10.1007/s11626-010-9343-x An improved method of cell culture system from eye stalk, hepatopancreas, muscle, ovary, and hemocytes of Penaeus vannamei [Elektronische Daten] [Sunil George, Arun Dhar] Improved methods of cell culture from eye stalk, hepatopancreas, muscle, ovary, and hemocytes of shrimp (Penaeus vannamei) were established using synthetic media and shrimp muscle extract (SME). For hemocytes and ovarian cell cultures, Grace's insect medium supplemented with 10% (v/v) fetal bovine serum and 10% SME (v/v) showed enhanced attachment and proliferation of the cells. The hemocyte and ovarian cell cultures could be maintained for 48 and 66days, respectively, and have been sub-cultured four and six times, respectively. Both ovary and hemocyte cell cultures contained primarily epithelial-like cells. Cells derived from ovary tissue grew preferably between 26°C and 28°C with 5% CO2. Although the temperature preference of hemocyte cells was the same as ovarian cells, CO2 supplementation did not show any difference in the growth of hemocyte cells. When the shrimp were injected with lipopolysaccharide (8μg/g of shrimp) and hemolymph was drawn 24h post-injection, the in vitro multiplicity of hemocytes dramatically improved. The growth of eye stalk, hepatopancreas, and muscle-derived cells was much less compared to ovarian cells and hemocytes under the conditions described above. The optimal culture conditions for ovarian cells and hemocytes were also different from that for eye stalk, hepatopancreas, and muscle cell culture. The proliferation efficiencies of primary cultures of hepatopancreas, eyestalk, and muscle cells were about 30, 12, and <7d, respectively. The improved culture conditions described here, particularly for hemocytes and ovary, will be very useful for in vitro studies involving viruses infecting shrimp and in shrimp genomic studies. The Society for In Vitro Biology, 2010 Penaeus vannamei nationallicence Primary cell culture nationallicence Eye stalk nationallicence Hepatopancreas nationallicence Muscle nationallicence Ovary nationallicence Hemocytes nationallicence George Sunil Advanced BioNutrition Corporation, 21046, Columbia, MD, USA aut Dhar Arun Advanced BioNutrition Corporation, 21046, Columbia, MD, USA aut In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/9(2010-10-01), 801-810 1071-2690 46:9<801 2010 46 11626 https://doi.org/10.1007/s11626-010-9343-x text/html Onlinezugriff via DOI 1 review-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11626-010-9343-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- George Sunil Advanced BioNutrition Corporation, 21046, Columbia, MD, USA aut NATIONALLICENCE 700 1- Dhar Arun Advanced BioNutrition Corporation, 21046, Columbia, MD, USA aut NATIONALLICENCE 773 0- In Vitro Cellular & Developmental Biology - Animal Springer-Verlag 46/9(2010-10-01), 801-810 1071-2690 46:9<801 2010 46 11626 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer