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The human diet contains a great variety of natural carcinogens, as well as many natural anticarcinogens. The physiological effects of food factors are of particular interest because they may be useful for human cancer prevention. Techniques for human cell culture will be useful both for basic studies of cell biology and for evaluation of physiologically active substance in vitro. Shellfishes are popular source of traditional Chinese medicine. In this study, Taiwan small abalone, Haliotis diversicolor was selected for immunological function assessment. Small abalone was separated into the viscera and muscle. The muscle was extracted by 40% ethanol, then boiled with hot-water (100°C, 30 min) and 50% ammonium sulfate saturation precipitation. In order to clarify performance at the cellular level, human-derived hybridoma and macrophage were cultured and treated in a serum-free system. The cell proliferation activation was used as indicators of function. Both hot-water extracts and ethanol extracts of small abalone promoted cell proliferation, immunoglobulin production and cell aggregation toward human hybridoma cell lines HB4C5, and promoted cell proliferation toward macrophage-like cell lines (U-937、U-M、THP-1 and J774.1). Small abalone hot-water extracts showed stable in acidic-base buffer treatment (pH 2-10). Cell proliferation activity remained about 80%, when being treated at 100oC for 60 min. In presence of small abalone hot-water extracts, the activity will be inhibited by N-Acetyl-D-glucosamine and maltose, these effectors on SAH and receptor interaction for cell activation are important. After purified by using Phenyl-TOYOPEARL hydrophobic interaction chromatography and DEAE anion-exchange chromatography, the proliferating factor was gained from SAH. The molecular weight of purified fraction was confirmed about 25-30 kDa by SDS-PAGE. In the absence of serum-free, the promoting factor of small abalone induced nitrite production after cultured J774.1 cell for 24, 48 and 72 h. In prescence of 5% serum, the promoting factor of small abalone induced nitrite production remarkably, and showed synergistic effect with IFN-g. The result obtained in this study indicated that the extract of small abalone should be good candidate for health food development. Further studies including structure, function and in vivo animals experiment of small abalone are needed for advanced health claim.
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