TY - JOUR
T1 - Hemolymph analysis and evaluation of newly formulated media for culture of shrimp cells (Penaeus Stylirostris)
AU - Shimizu, Chisato
AU - Shike, Hiroko
AU - Klimpel, Kurt R.
AU - Burns, Jane C.
N1 - Funding Information:
This project was funded in part by Super Shrimp Inc. and by a grant from the National Sea Grant College Program, National Oceanic and Atmospheric Administration, U.S. Department of Commerce, under grant NA66RG0477, project R/A-113, through the California Sea Grant College system. This pro-jeet was performed with technical support from Richard Gattra, MT, CLS, MBA; Senior Clinical Laboratory Specialist, UCSD Medical Center; Herman van Halbeek, Ph.D., Glycotechnology Core Resource of the UCSD Glyeo-biology Research and Training Center; Bruce A. Barshop, M.D., Division of Biochemical Genetics, UCSD School of Medicine; and Kevin Walda, Ph.D., Analytical Facility, Scripps Institute of Oceanography. We thank Tony Dettori and the support staff at Super Shrimp Inc. for technical assistance.
PY - 2001
Y1 - 2001
N2 - Creation of a shrimp cell line has been an elusive goal. This failure may be due to the composition of the cell culture medium, which may be inadequate to support primary cultured cells. Shrimp hemolymph should contain the nutritional components needed to support cell growth and division. We report here the comprehensive biochemical analysis of hemolymph from the blue shrimp, Penaeus stylirostris (Litopenaeus stylirostris) (see Holthuis, L. B. Shrimps and prawns of the world, in: FAO species catalog. Vol. 1. Rome: Food and Agriculture Organization of the United Nations; 1980), for free amino acids (FAAs), carbohydrates, electrolytes, metals, pH, and osmolality. Levels of hemolymph components were compared to 2×L-15 with 20% fetal bovine serum, a commonly used culture medium for crustacean cells. The FAAs, taurine and proline, and the metals, strontium and zinc, were significantly higher in hemolymph than in the 2×L-15 medium. In contrast, other FAAs were up to 50 times higher in the 2×L-15 medium than in the hemolymph. To mimic more closely the hemolymph composition, we created two new media based on either the 0.2×L-15 or the M199 medium. We compared the microscopic appearance of cells cultured in these media and evaluated deoxyribonucleic: Acid (DNA) and protein synthesis by 3H-thymidine uptake and 3S-methionine uptake assays. The ovary cells of P. stylirostris cultured in either of the new media formed monolayers, while the cells cultured in 2×L-15 medium did not. Despite these differences, there was no evidence of sustained DNA or protein synthesis with any of the media. Future studies to establish a shrimp cell line should focus on analysis of the cell cycle and on overcoming the molecular blocks to cell division.
AB - Creation of a shrimp cell line has been an elusive goal. This failure may be due to the composition of the cell culture medium, which may be inadequate to support primary cultured cells. Shrimp hemolymph should contain the nutritional components needed to support cell growth and division. We report here the comprehensive biochemical analysis of hemolymph from the blue shrimp, Penaeus stylirostris (Litopenaeus stylirostris) (see Holthuis, L. B. Shrimps and prawns of the world, in: FAO species catalog. Vol. 1. Rome: Food and Agriculture Organization of the United Nations; 1980), for free amino acids (FAAs), carbohydrates, electrolytes, metals, pH, and osmolality. Levels of hemolymph components were compared to 2×L-15 with 20% fetal bovine serum, a commonly used culture medium for crustacean cells. The FAAs, taurine and proline, and the metals, strontium and zinc, were significantly higher in hemolymph than in the 2×L-15 medium. In contrast, other FAAs were up to 50 times higher in the 2×L-15 medium than in the hemolymph. To mimic more closely the hemolymph composition, we created two new media based on either the 0.2×L-15 or the M199 medium. We compared the microscopic appearance of cells cultured in these media and evaluated deoxyribonucleic: Acid (DNA) and protein synthesis by 3H-thymidine uptake and 3S-methionine uptake assays. The ovary cells of P. stylirostris cultured in either of the new media formed monolayers, while the cells cultured in 2×L-15 medium did not. Despite these differences, there was no evidence of sustained DNA or protein synthesis with any of the media. Future studies to establish a shrimp cell line should focus on analysis of the cell cycle and on overcoming the molecular blocks to cell division.
UR - http://www.scopus.com/inward/record.url?scp=0034877270&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034877270&partnerID=8YFLogxK
U2 - 10.1290/1071-2690(2001)037<0322:HAAEON>2.0.CO;2
DO - 10.1290/1071-2690(2001)037<0322:HAAEON>2.0.CO;2
M3 - Article
C2 - 11515962
AN - SCOPUS:0034877270
SN - 1071-2690
VL - 37
SP - 322
EP - 329
JO - In Vitro Cellular and Developmental Biology - Animal
JF - In Vitro Cellular and Developmental Biology - Animal
IS - 6
ER -