Bacterial adhesion and invasion assay

hCMEC/D3 cells (obtained from Millipore) were grown in EndoGRO-MV complete media (Millipore, SCME004) supplemented with 5% fetal bovine serum and fibroblast growth factor 2 (1 ng/ml; Millipore). Cells were grown in tissue culture treated 24-well plates and 5% CO₂ at 37°C. Assays to determine the total number of bacteria adhered to host cells or intracellular bacteria were performed using GBS empty vector and complement strain as described previously (24 (link), 31 (link), 32 (link)). See Supplementary Text for more details.

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Joyce L.R., Kim S., Spencer B.L., Christensen P.M., Palmer K.L., Guan Z., Siegenthaler J.A, & Doran K.S. (2024). Streptococcus agalactiae glycolipids promote virulence by thwarting immune cell clearance. Science Advances, 10(22), eadn7848.

Publication 2024

hCMEC/D3 cells EndoGRO-MV complete media

Corresponding Organization : University of Colorado Denver

Other organizations : The University of Texas at Dallas, Duke University Hospital, Duke Medical Center

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Variable analysis

independent variables

GBS empty vector
GBS complement strain

dependent variables

Total number of bacteria adhered to host cells
Number of intracellular bacteria

control variables

HCMEC/D3 cells (obtained from Millipore)
EndoGRO-MV complete media (Millipore, SCME004) supplemented with 5% fetal bovine serum and fibroblast growth factor 2 (1 ng/ml; Millipore)
Tissue culture treated 24-well plates
5% CO2 at 37°C

controls

Positive control: GBS complement strain
Negative control: GBS empty vector

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