Thanks to Dr. David Pennock at Miami University (Oxford, OH) I am able to offer four culture medias for use with Tetrahymena.

1. Starvation medium:

10 mM Tris, pH 7.4 ('nuff said)

2. Neff's Growth medium

 Reagent

 grams per liter

 0.1 mM Ferric Citrate

 0.024 g

 2.0 mM KH2PO4

 0.27 g

 0.05 mM CaCl2

 0.007 g

 1.0 mM MgSO4-7H20

  0.25 g

 0.5% Glucose

 5.0 g

0.25% yeast extract  

 2.5 g

 0.25% proteose peptone

  2.5 g

Procedure:

1. Dissolve ferric citrate in 1/4 volume of boiling water.

2. Once the ferric citrate is dissolved, add water to 3/4 volume.

3. Dissolve the remaining ingredients in the order listed above.

4. Bring to volume, aliquot, autoclave.

This medium is good for growing cells for genetics. Cells will mate after being diluted 1:5 with 10 mM Tris, pH 7.4. Cells have to be spun down and washed when grown in other media.

 

More below.

 
 

 

3. Super Proteose Peptone Growth medium

Reagent

 grams per liter

 1% proteose peptone

 10 g

 0.2% glucose

 2 g

 0.1% yeast extract

 1 g

 0.003% EDTA (ferric salt)

 0.03 g

Dissolve all ingredients in 3/4 volume of water. Bring to volume, aliquot, autoclave.

Comments: Dr. Penncok uses Difco proteose peptone and yeast extract. Cells are grown with shaking at 125 - 150 rpm at 28°. Cells are grown in Erlenmeyer flasks, filled approximately 1/5 to 1/4 full, i.e. put 50 - 60 ml in a 250 ml flask, 125 ml in a 500 ml flask and 250 ml in a 1 l flask.

 

4. Cell Maintenance medium

 Reagent

 grams per liter

 2% proteose peptone

 20 g

 0.1 % yeast extract

 1 g

 0.2% glucose

 2 g

Procedure:

1. Dissolve all ingredients in 3/4 volume water.

2. Bring to volume. Aliquot 6 ml/tube. Autoclave.

Comments: Transfer cells every 3 - 4 weeks by moving 0.5 to 1.0 ml from the old test tube to a new test tube.
Last updated Wednesday July 4, 2001 Webmaster Dean Fraga.