Summary
We describe a rapid method for the isolation of large numbers of livingHydra cells of defined cell type in an isotonic cell medium (Gierer et al. 1972). Intact animals are enzymatically dissociated into a single cell suspension and the various cell types separated in less than one hour by counterflow centrifugation elutriation. Cell loss is minimal. RNA isolated from various fractions can be probed with cell type specific cDNA-clones.
Similar content being viewed by others
References
Bode HR, Gee LW, Chow M (1990) Neuron differentiation in hydra involves dividing intermediates. Dev Biol 139:231–243
Bode H, Berking S, David CN, Gierer A, Schaller H, Trenkner E (1973) Quantitative analysis of cell types during growth and morphogenesis inHydra. Roux' Arch Dev Biol 171:269–285
Chomczynski P, Sacchi N (1987) Single-Step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Conkie D (1986) Separation of viable cells by centrifugal elutriation. In: Freshney RI (ed) Animal cell culture. Oxford IRL Press, Oxford, pp 113–134
David CN (1973) A quantitative method for maceration ofHydra tissue. Roux' Arch Dev Biol 171:259–268
David CN, Bosch TCG, Hobmayer B, Holstein ThW, Schmidt T (1987) Interstitial stem cell in hydra. In: Loomis WF (ed) Genetic regulation of development. AR Liss, New York, pp 389–408
Gierer A, Berking S, Bode H, David CN, Flick K, Hausmann G, Schaller H, Trenkner E (1972) Regeneration of Hydra from reaggregated cells. Nature 239:98–105
Hayles J, Aves S, Nurse P (1986)Suc1 is an essential gene involved in both the cell cycle and growth in fision yeast. EMBO J 5:3373–3379
Holstein ThW (1981) The morphogenesis of nematocytes inHydra and Forskalia: an ultrastructural study. J Ultrastruct Res 75:276–290
Holstein ThW, David CN (1990a) Cell cycle length, cell size, and proliferation rate in hydra stem cells. Dev Biol 142:392–400
Holstein ThW, David CN (1990b) Putative intermediates in the nerve cell differentiation pathway in hydra have properties of multipotent stem cells. Dev Biol 142:401–405
Kurz E, Holstein ThW, Petri BM, Engel J, David CN (1991) Minicollagens inHydra nematocytes. J Cell Biol 115:1159–1169
MacWilliams HK (1991) Models of pattern formation.Hydra andDictyostelium. Semin Dev Biol 2:119–128
Lindahl PE (1948) Principle of a counter-streaming centrifuge for separation of particles of different sizes. Nature 161:648–649
Sambrook J, Fritsch EF, Maniatis Th (1989) Molecular cloning. A laboratory manual. (2nd edition) Cold Spring Harbor Laboratory Press, New York
Schaller H, Gierer A (1973) Distribution of the head-activating substance in hydra and its localisation in membranous particles in nerve cells. J Embryol Exp Morphol 29:39–52
Schartl M, Holstein ThW, Robertson SM, Barnekow A (1989) Preferential expression of a pp60c-src related tyrosine kinase activity in nerve cells of the early metazoanHydra (Coelenterates). Oncogene 4:1185–1191
Stei A (1988) Entwicklung von Methoden zur Verwendung von Hydrazellen im Durchflußzytometer. Diploma-Thesis, Zoologisches Institut der Ludwig-Maximilians-Universität München: S 1–75
Sugiyama T, Fujisawa T (1977) Genetic analysis of developmental mechanisms inHydra. I. Sexual reproduction ofHydra magnipapillata and isolation of mutants. Dev Growth Differ 19:187–200
Technau U, Holstein ThW (1992). Cell sorting during the regeneration of Hydra from reaggreated cells. Dev Biol 151 (in press).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Greber, M.J., David, C.N. & Holstein, T.W. A quantitative method for separation of livingHydra cells. Roux's Arch Dev Biol 201, 296–300 (1992). https://doi.org/10.1007/BF00592110
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00592110