Hemolymph analysis and evaluation of newly formulated media for culture of shrimp cells (Penaeus Stylirostris)

Chisato Shimizu, Hiroko Shike, Kurt R. Klimpel, Jane C. Burns

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)322-329
Number of pages8
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume37
Issue number6
DOIs
StatePublished - Aug 29 2001

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology

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