Abstract
In order to acquire more in-depth information on the site(s) of hemopoiesis in marine animals, we successfully cultured pharyngeal explants of the tunicate Styela clava over a period of 82 d. Transmission electron microscopy of resident hemocytes within explants revealed changes in hemocyte composition. Hemoblast-like cells increased shortly after commencement of the cultures (8.1% at Day 0 increasing to a maximum of 28.7% at Day 7). Autoradiography using 3H-thymidine incorporation confirmed that hemocyte proliferation in pharyngeal explants still continued after 37 d culture. During culture, the migration of many free cells into the medium resulted in sparse, residential hemocytes in the pharyngeal explants. Hemocyte migration increased by up to 4.3x105 cells/explant (max.) at 17 to 24 d, but finally decreased to 4.9x104 cells/explant at 75 to 82 d. Vital neutral-red staining revealed that many emerging cells were not hemocytes such as those found in the normal hemolymph. The continued development of in vitro approaches will strengthen analyses of immune-defense responses in tunicates which, as protochordates, are the immediate invertebrate ancestors of vertebrates.
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References
Azumi K, Yokozawa H, Ishii S (1991) Lipopolysaccharide induces release of a metallo-protease from hemocytes of the ascidian, Halocynthia roretzi. Devl comp Immun, NY 15:1–7
Beck G, O'Brien RF, Habicht GS, Stillman, DL, Cooper EL, Raftos DA (1993) Invertebrate cytokines. III: Invertebrate interleukin-1-like molecules stimulate phagocytosis by tunicate and echinoderm cells. Cell Immun 146:284–299
Beck G, Vasta G, Marchalonis JJ, Habicht GS (1989) Characterization of interleukin-1 activity in tunicates. Comp Biochem Physiol 92 B:93–98
Cooper EL, Rinkevich B, Uhlenbruck G, Valembois P (1992) Invertebrate immunity: another viewpoint. Scand J Immun 35:247–266
Ermak TH (1975) An autoradiographic demonstration of blood cell renewal in Styela clava (Urochordata: Ascidiacea). Experientia 31:837–839
Ermak TH (1976) The hematogenic tissues of tunicates. In: Wright RK, Cooper EL (eds) Phylogeny of thymus and bone marrowbursa cells. Elsevier/North-Holland, Amsterdam, pp 45–56
Fuke MT (1980) “Contact reactions” between xenogeneic or allogeneic coelomic cells of solitary ascidians. Biol Bull mar biol Lab, Woods Hole 158:304–315
Kelly KL, Cooper EL, Raftos DA (1992) Purification and characterization of a humoral opsonin from the solitary urochordate Styela clava. Comp Biochem Physiol 103 B:749–753
Kelly KL, Cooper EL, Raftos DA (1993a) A humoral opsonin from the solitary urochordate Styela clava. Devl comp Immun, NY 17:29–39
Kelly KL, Cooper, EL, Raftos DA (1993b) Cytokine-like activities of a humoral opsonin from the solitary urochordate Styela clava. Zool Sci 10:57–64
Raftos DA, Cooper EL, Habicht GS, Beck G (1991) Invertebrate cytokines-tunicate cell proliferation stimulated by an interleukin-1-like molecule. Proc natn Acad Sci USA. 88:9518–9522
Raftos DA, Cooper EL, Stillman DL, Habicht GS, Beck G (1992) Invertebrate Cytokines II: release of interleukin-1-like molecules from turicate hemocytes stimulated with zymosan. Lymphokine Cytokine Res 11:235–240
Raftos DA, Stillman DL, Cooper EL (1990) In vitro culture of tissue from the tunicate Styela clava. In vitro Cell Dev Biol 26:962–970
Rowley AF (1981) The blood cells of the sea squirt, Ciona intestinalis: morphology, differential counts and in vitro phagocytic activity. J Invertebr Path 37:91–200
Rowley AF (1982) Ultrastructural and cytochemical studies on the blood cells of the sea squirt, Ciona intestinalis. I. Stem cells and amoebocytes. Cell Tissue Res 223:403–414
Sawada T, Fujikura Y, Tomonaga S, Fukumoto T (1991) Classification and characterization of ten types of hemocytes in tunicate Halocynthia roretzi. Zool Sci 8:939–950
Sawada T, Zhang J, Cooper EL (1993) Classification and characterization of hemocytes of tunicate Styela clava. Biol Bull mar biol Lab, Woods Hole 184:87–96
Watanabe H, Taneda Y (1982) Self or non-self recognition in compound ascidians. Am Zool 22:775–782
Wright RK (1981) Urochordates. In: Ratcliffe, Rowley (eds) Invertebrate blood cells. Vol. 2. Academic Press, London, pp 565–626
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Communicated by M. G. Hadfield, Honolulu
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Sawada, T., Zhang, J. & Cooper, E.L. Sustained viability and profileration of hemocytes from the cultured pharynx of Styela clava . Marine Biology 119, 597–603 (1994). https://doi.org/10.1007/BF00354323
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DOI: https://doi.org/10.1007/BF00354323