Adapted from Hirumi & Hirumi (1994) Parasitology Today 10:80
HMI-9 powder is custom made by Life Technologies and is generally purchased on a consortium basis
One pot is sufficient to make up 5 litres after addition of the following:
|Sodium bicarbonate||15 g|
|Pen-Strep (Life Technologies)||50 ml|
Make up to 4.5 litres with distilled water, filter sterilise, store at 4°C in 450 ml volumes.
Prior to use, add 50 ml fetal bovine serum (FBS; Sigma), giving a final concentration of 10% FBS and store at 4°C for up to two months.
NB: Some early publications indicate that FBS should be heat inactivated at 56°C for 30 minutes prior to use in bloodstream-form T. brucei culture – this is not necessary; in fact our unpublished data suggests that using non-inactivated FBS leads to faster, more robust cell growth.
Cultures should be grown in 5% CO2 incubator at 37°C, and should not exceed a density of 2.5×106 cells per ml.