Well cell growth or toxicity assays

PRELIMINARY PROCEDURE 96-WELL CELL GROWTH OR TOXICITY ASSAYS 

Using 8-well multichannel pipettes, plate the cell suspensions (carefully mixed) into 96-well tissue culture dishes (e.g. Costar or Greiner). 

Incubate at 37°C in a humidified 95% air/5% CO2 atmosphere. Total volume of growth medium per well should be 100 p.1. 

After an appropriate growth period (e.g. 4-6 days, depending on cell line, for serum batch testing), determine the endpoint (as below). For toxicity tests, incubate cells in 100 xl of medium for 24 h prior to addition of test substance (at twice the final concentration in 100 xl of medium). Incubation with toxic substance prior to endpoint measurement is usually for 6 days. For measurement of Trypan blue exclusion, trypsinize cells with 0.5% trypsin (Gibco) and 

Trypan blue is harmful if ingested or inhaled. It is irritating to the eyes, is harmful by skin contact and has been found to cause cancer in laboratory animals. Appropriate precautions should be taken when handling Trypan blue and the use of an extraction hood and gloves is advised if handling Trypan blue in powder form. 

---