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1. Harvest cells from a culture which is at or near peak density by centrifugation at ~800 x g for 5 min.
2. Adjust concentration of cells to 0.5–1.0 x 10⁷/mL in fresh growth medium.  If the concentration is too low, centrifuge at ~800 x g for 5 minutes and resuspend the cell pellet with a volume of supernatant to yield the desired concentration.
3. While cells are centrifuging, prepare a 20% (v/v) solution of sterile glycerol in fresh growth medium. 
4. Mix the cell preparation and the glycerol solution in equal portions. The final concentration will be 2.5-5 x 10⁶ cells/mL in 10% glycerol. The time from the mixing of the cell preparation and glycerol stock solution before the freezing process is begun should be no less than 15 min and no more than 30 min.
5. Dispense in 0.5 mL aliquots into 1.0 - 2.0 mL sterile plastic screw-capped cryovials.
6. Place the ampules in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)  If freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through heat of fusion.  At -40ºC, plunge ampules into liquid nitrogen.
7. Store in either the vapor or liquid phase of a nitrogen refrigerator.
8. To thaw a frozen ampule, place it in a 35°C water bath such that the lip of the ampule remains above the water line. Thawing time is approximately 2 to 3 minutes.  Do not agitate the ampule.  Do not leave ampule in water bath after it is thawed.
9. Remove the vial from the water bath immediately after thawing.  Aseptically transfer the contents of the ampule into 10 mL of fresh growth medium. 
10. Incubate the flask at 27°C with the cap screwed on tightly.
11. Maintain as described above. 

Goedbloed E, et al. Serological studies of trypanosomiasis in East Africa. II. Comparisons of antigenic types of Trypanosoma brucei subgroup organisms isolated from wild tsetse flies. Ann Trop med Parasitol 67(1): 31-43, 1973. PubMed: 4723213

Gibson W. The origins of the trypanosome genome strains Trypanosoma brucei brucei TREU 927, T. b. gambiense DAL 972, T. vivax Y486 and T. congolense IL3000. Parasit Vectors 5: 71, 2012. PubMed: 22483376

van Deursen FJ, et al. Characterisation of the growth and differentiation in vivo and in vitro-of bloodstream-form Trypanosoma brucei strain TREU 927. Mol BIochem Parasitol 112(2): 163-171, 2001. PubMed: 11223123

Berriman M, et al. The genome of the African trypanosome Trypanosoma brucei. Science 309(5733): 416-422, 2005. PubMed: 16020726

Nucleotide (GenBank) : AL929603 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 1, complete sequence

Nucleotide (GenBank) : AE017150 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 2, complete sequence

Nucleotide (GenBank) : CP000066 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 3, complete sequence

Nucleotide (GenBank) : CP000067 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 4, complete sequence

Nucleotide (GenBank) : CP000068 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 5, complete sequence

Nucleotide (GenBank) : CP000069 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 6, complete sequence

Nucleotide (GenBank) : CP000070 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 7, complete sequence

Nucleotide (GenBank) : CP000071 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 8, complete sequence

Nucleotide (GenBank) : CM000207 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 9, whole genome shotgun sequence

Nucleotide (GenBank) : CM000208 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 10, whole genome shotgun sequence

Nucleotide (GenBank) : CH464492 Trypanosoma brucei brucei strain 927/4 GUTat10.1 chromosome 11 chr11_scaffold02 genomic scaffold, whole genome shotgun sequence