Cultivation of Uninfected and A. phagocytophilum-Infected Human and Macaque Cells

Uninfected and A. phagocytophilum NCH-1 strain infected human promyelocytic HL-60 cells (ATCC CCL-240; American Type Culture Collection [ATCC]) were cultured as previously described (30 (link)). Uninfected and infected RF/6A Macaca mulatta choroidal endothelial cells (CRL-1780; ATCC) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with l-glutamine, 4.5 g d-glucose and 100 mg sodium pyruvate (Gibco) supplemented with 10% (vol/vol) heat-inactivated fetal bovine serum (FBS) (Gemini Bioproducts), 1X minimal essential medium containing non-essential amino acids (Gibco) and 15 mM HEPES (Gibco) in a humidified incubator at 37°C with 5% atmospheric CO₂.

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Read C.B., Lind M.C., Chiarelli T.J., Izac J.R., Adcox H.E., Marconi R.T, & Carlyon J.A. (2022). The Obligate Intracellular Bacterial Pathogen Anaplasma phagocytophilum Exploits Host Cell Multivesicular Body Biogenesis for Proliferation and Dissemination. mBio, 13(6), e02961-22.

Publication 2022

CCL-240 CRL-1780 l-glutamine d-glucose sodium pyruvate fetal bovine serum (FBS) non-essential amino acids HEPES

Choroidal Eagle Endothelial cells Essential amino acids Fetal bovine serum Glucose Hepes Hl 60 cells Human L glutamine Macaca mulatta Promyelocytic Pyruvate Sodium Strain

Corresponding Organization : Virginia Commonwealth University Medical Center

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Variable analysis

independent variables

Uninfected and A. phagocytophilum NCH-1 strain infected human promyelocytic HL-60 cells
Uninfected and infected RF/6A Macaca mulatta choroidal endothelial cells

dependent variables

Not explicitly mentioned

control variables

Cell culture conditions: Dulbecco's modified Eagle's medium (DMEM) with l-glutamine, 4.5 g d-glucose and 100 mg sodium pyruvate, supplemented with 10% (vol/vol) heat-inactivated fetal bovine serum (FBS), 1X minimal essential medium containing non-essential amino acids, and 15 mM HEPES, in a humidified incubator at 37°C with 5% atmospheric CO2

controls

Uninfected cells as a negative control

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