Culturing JEG-3 Cells and HBMECs

2D-cultured JEG-3 cells (2D SYNs; ATCC® HTB-36™, American Type Culture Collection, VA), human brain microvascular endothelial cells [HBMECs; (Coyne et al., 2007 (link))], LLC-MK2 cells (ATCC® CCL-7™; American Type Culture Collection, VA), and three-dimensional (3D) cultures of JEG-3 cells (3D SYNs) were grown as described previously (Silberstein et al., 2021 (link)). Experiments with 3D SYNs and 3D HBMECs were carried out between day 20–22 and day 4–6 after culture initiation, respectively. The number of viable cells was determined using the CellTiter-Fluor™ (for 2D SYNs) or the CellTiter-Glo™ 3D (for 3D SYNs) cell viability assays following manufacturer’s instructions (Promega, WI).

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Silberstein E., Chung C.C, & Debrabant A. (2023). The transcriptome landscape of 3D-cultured placental trophoblasts reveals activation of TLR2 and TLR3/7 in response to low Trypanosoma cruzi parasite exposure. Frontiers in Microbiology, 14, 1256385.

Publication 2023

ATCC® CCL-7™; CellTiter-Fluor™ CellTiter-Glo™ 3D

Assays Brain Cell viability Cells Cells cultures Cultured cells Endothelial cells Human Promega

Corresponding Organization : Center for Biologics Evaluation and Research

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Variable analysis

independent variables

2D-cultured JEG-3 cells (2D SYNs)
Human brain microvascular endothelial cells [HBMECs]
LLC-MK2 cells
Three-dimensional (3D) cultures of JEG-3 cells (3D SYNs)

dependent variables

Number of viable cells

control variables

Culture conditions for 2D SYNs, 3D SYNs, and 3D HBMECs as described previously (Silberstein et al., 2021)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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