Cultivation of Procyclic and Bloodstream Trypanosoma brucei

Procyclic forms of T. brucei strain Lister 427 were cultivated at 27 °C in SDM-79 media [52 (link)], supplemented with 10% heat inactivated fetal bovine serum (Sigma), 0.2% (v/v) of hemin (Sigma) solution at 2 mg mL⁻¹ in 0.2 M NaOH, and 1% (v/v) penicillin and streptomycin (Sigma). Bloodstream forms of T. brucei strain Lister 427 were maintained at 37 °C in a humidified incubator with 5% CO₂ in HMI-9 media [53 (link)], supplemented with 10% heat inactivated fetal bovine serum (Sigma) and 1% (v/v) penicillin and streptomycin (Sigma). CSB RNAi knockdown and Rad51 knockout BSF strains were generated and described previously in [29 (link)] and [51 (link)].

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Vieira-da-Rocha J.P., Passos-Silva D.G., Mendes I.C., Rocha E.A., Gomes D.A., Machado C.R, & McCulloch R. (2019). The DNA damage response is developmentally regulated in the African trypanosome. DNA Repair, 73, 78-90.

Publication 2019

fetal bovine serum hemin penicillin and streptomycin

Bloodstream Fetal bovine serum Hemin Penicillin Rnai Strains Streptomycin

Corresponding Organization : Wellcome Centre for Molecular Parasitology

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Variable analysis

independent variables

Cultivation temperature (27 °C for procyclic forms, 37 °C for bloodstream forms)
Media (SDM-79 for procyclic forms, HMI-9 for bloodstream forms)
Cell lines (Procyclic forms of T. brucei strain Lister 427, Bloodstream forms of T. brucei strain Lister 427, CSB RNAi knockdown BSF strain, Rad51 knockout BSF strain)

dependent variables

Not explicitly mentioned

control variables

Supplementation of media (10% heat inactivated fetal bovine serum, 0.2% (v/v) of hemin solution, 1% (v/v) penicillin and streptomycin)
Incubation conditions (5% CO2 for bloodstream forms)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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