Tetrahymena Cell Culture


Tetrahymena, a free‐living unicellular eukaryote, is a ciliate considered to be the most highly developed protozoan, because it possesses specialised organelles that perform each of the cell functions. This pear‐shaped microorganism (30×50 μm) can be found in almost all freshwater environments, including even natural hot springs in which a thermotolerant strain can live. One distinct feature that sets ciliate apart from other multicellular eukaryotes is the possession of two functionally different nuclei – macronucleus, which is transcriptionally active and responsive for the somatic function of cells, and micronucleus, which is transcriptionally silent and serves as germinal nucleus during mating. Tetrahymena grows easily to high density. Its relatively large size, nuclear dimorphism and other unique genetic characteristics make it a good model system to study the molecular, cellular and genetic processes of cells.

Key Concepts:

  • Ciliates are unicellular organisms that execute many biological activities observed in multicellular organisms.

  • Tetrahymena cells can survive under extremely dangerous conditions such as cold, pH alterations and osmotic stress.

  • Tetrahymena cells possess rapid adaptation machineries to regulate biological processes including cell division, conjugation and cell death against intracellular and extracellular stresses.

Keywords: protozoa; Tetrahymena; cell culture; synchrony; temperature shift; starvation; conjugation

Figure 1.

The synchronous cell division of Tetrahymena induced by heat treatment. Tetrahymena pyriformis strain w cells were subjected to eight cycles of heat treatment as described in the text and harvested at the indicated time points. Cells were fixed with 2% formaldehyde and stained with Hoechst 33258. Images were taken under fluorescent microscope. EHT, end of heat treatment.

Figure 2.

Conjugation of Tetrahymena. Tetrahymena thermophila strain CU428 and B2086 were prestarved for 24 h and then mixed together equally. Cells were harvest at 3 h (a) and 7 h (b) after mixing and stained with (DAPI). White arrows indicate macronuclei and red arrows indicate micronuclei.



Kimura M, Hirono M, Takemasa T and Watanabe Y (1991) Drastic change in the level of actin mRNA in the course of synchronous division in Tetrahymena. Journal of Biochemistry 109(3): 399–403.

Nakashima S, Wang S, Hisamoto N et al. (1998) Molecular cloning and expression of a stress‐responsive mitogen‐activated protein kinase‐related kinase from Tetrahymena cells. Journal of Biological Chemistry 274(15): 9976–9983.

Raikov IB (1972) Nuclear phenomena during conjugation and autogamy in ciliates. In: Chen TT (ed.) Research in Protozoology, vol. 4, pp. 147–290. Elmsford, NY: Pergamon.

Thompson GA Jr and Nozawa Y (1977) Tetrahymena: a system for studying dynamic membrane alterations within the eukaryotic cell. Biochimica et Biophysica Acta 472(1): 55–92.

Wang S, Nakashima S, Numata O, Fujiu K and Nozawa Y (1999) Molecular cloning and cell‐cycle‐dependent expression of the acetyl‐CoA synthetase gene in Tetrahymena cells. Biochemical Journal 343: 479–485.

Wang S, Nakashima S, Sakai H et al. (1998) Molecular cloning and cell‐cycle‐dependent expression of a novel NIMA (never‐in‐mitosis in Aspergillus nidulans)‐related protein kinase (TpNrk) in Tetrahymena cells. Biochemical Journal 334: 197–203.

Further Reading

Elliott AM (ed.) (1973) The Biology of Tetrahymena. Stroudsburg, PA: Dowden, Hutchinson and Ross.

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Zhang, Siwei, Ling, Zihui, Wang, Shulin, Nozawa, Yoshinori, and Umeki, Shigenobu(Sep 2010) Tetrahymena Cell Culture. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002576.pub2]