Culture of suspension cells in a spinner flask

PROCEDURE CULTURE OF SUSPENSION CELLS IN A SPINNER FLASK 

Standard tissue culture media can be used with various supplements. Serum at 10% can cause foaming at fast stirring speeds, so the serum level is usually reduced to as low a level as possible (0.5-5%). The use of HEPES buffer to stabilize the pH during the setting-up procedure is beneficial. 

Cell and Tissue Culture Laboratory Procedures in Biotechnology, edited by A. Doyle and J.B. Griffiths. 1998 John Wiley Sons Ltd. 

Pluronic F-68 (polyglycol) (BASF, Wyandot) can be used at 0.1% to protect cells against mechanical damage, especially at reduced serum concentrations. Carboxymethyl cellulose (CMC) (15-20 cP) can also be used to protect cells from mechanical damage at 0.1% concentration. Antifoam (6 ppm) (Dow Chemical Co.) is recommended at serum concentrations above 2-3%. For more information, see Chapter 4, section 4.8. 

Add cells from a logarithmically growing seed culture at 1-2 x 10 cells mH. 

---