Development and characterization of primary cell cultures from the hematopoietic tissues of the Dublin Bay prawn, Nephrops norvegicus

A. L. Mulford, F. Lyng, C. Mothersill, B. Austin

    Research output: Contribution to journalArticle

    22 Citations (Scopus)

    Abstract

    Improved maintenance in vitro of the hematopoietic tissue of the Dublin Bay prawn Nephrops norvegicus (L.) resulted by using 10% (v/v) 2× Leibovitz's medium prepared in seawater (salinity = 25‰), and supplemented with 10% (v/v) heat inactivated fetal bovine serum plus 5% (v/v) Nephrops serum or 5% (v/v) Nephrops muscle extract, and 0.06 g/l of L-proline and 1 g/l of glucose. Pronase at 100 µg/ml improved tissue dissociation and subsequent spreading of hematopoeitic cell cultures. The addition of epithelial growth factor (EGF), basic fibroblast growth factor (bFGF) or insulin growth factor 1 (IGF-1) did not enhance cell growth. Cell cultures contained several types of maturing hemocytes, in the size range of 6-24 µm diameter. Acid phosphatase, a-naphthyl butyrate esterase, a-naphthyl acetate esterase, naphthyl ASD chloroacetate esterase activity and phenoloxidase activity was demonstrated, but not so alkaline phosphatase or peroxidase. Small PAS (= Periodic acid Schiff) positive granules, unsaturated lipids and phospholipids were observed. Cultures remained functional for over two weeks. Mitosis was noticed occasionally; however, cell proliferation was not recorded by use of nuclear proliferation markers.

    Original languageEnglish
    Pages (from-to)265-275
    Number of pages11
    JournalMethods in Cell Science
    Volume22
    Issue number4
    DOIs
    Publication statusPublished - 2000

    Keywords

    • Hematopoietic tissue
    • Nephrops norvegicus
    • Primary cell cultures

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