Bme vitamins basal medium eagle

Laboratory colonies of Sergentomyia schwetzi (from Ethiopia), Phlebotomus orientalis (from Ethiopia), P. argentipes (from India) and P. papatasi (from Turkey) were maintained in the insectary of the Charles University in Prague under standard conditions (at 26 °C fed on 50% sucrose with a 14 h light/10 h dark photoperiod) as described previously [19 (link)]. Leishmania donovani (MHOM/ET/2010/GR374) transfected with green fluorescent protein (GFP) were cultured in M199 medium (Sigma) containing 10% heat-inactivated fetal bovine serum (FBS, Gibco) supplemented with 1% BME vitamins (Basal Medium Eagle, Sigma), 2% sterile urine, 250 μg/mL amikacin (Amikin, Bristol-Myers Squibb) and 150 μg/mL selective antibiotic G418 (Sigma). Leishmania major LV561 (LRC-L137; MHOM/IL/1967/Jericho-II) transfected with GFP protein were cultured in the same medium without selective antibiotics.

A laboratory colony of P. argentipes (originating from India) was maintained in the insectary of the Charles University in Prague under standard conditions (at 26 °C fed on 50% sucrose with a 14 h light/10 h dark photoperiod) as described previously (Volf and Volfova, 2011 (link)).
Amastigote Leishmania stages were grown in BMMs differentiated from precursor cells of BALB/c mice in the presence of L929 fibroblast cell culture supernatant as a source of macrophage-colony stimulating factor (M-CSF). Leishmania donovani (MHOM/ET/2010/GR374) promastigotes transfected with green fluorescence protein as described in Sadlova et al. (2011 (link)) were cultured in M199 medium (Sigma) containing 10% heat-inactivated fetal calf serum (Gibson) supplemented with 1% BME vitamins (Basal Medium Eagle, Sigma), 2% sterile urine, 250 µg mL⁻¹ amikacin (Amikin, Bristol-Myers Squibb) and 150 µg mL⁻¹ selective antibiotic G 418 (Sigma). Macrophages were exposed to stationary-phase parasites at a parasite-to-macrophage ratio of eight promastigotes to one macrophage. Both infected and uninfected macrophages were cultured in complete RPMI-1640 medium (Sigma) containing 10% FBS (fetal bovine serum), 20% L929 cell culture supernatant, 1% penicillin–streptomycin (Sigma), 2 mm of L-glutamine (Sigma) and 0·05 mm of β-mercapto-ethanol at 37 °C with 5% CO_2.