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Bradyzoite activation for infection of human intestinal microphysiological systems to study bradyzoite interactions.

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posted on 2025-02-04, 18:40 authored by Carlos J. Ramírez-Flores, Nicole D. Hryckowian, Andrew N. Gale, Kehinde Adebayo Babatunde, Marcos Lares, David J. Beebe, Sheena C. Kerr, Laura J. Knoll

(A) Schematic representation of bradyzoite infection niche in human intestinal MPS. (i) Live staining of actin showing the coverage of the lumen by Caco-2 cells. (ii) FITC-dextran live imaging assessing the polarity of the Caco-2 cell line in the MPS. (iii) Parasite replication at 48 hours post-infection with 1 × 103 RH tachyzoites as a control for host cell viability. (B) Representative fluorescent images of parasite replication in HFF cells infected with brain bradyzoites, either activated by pepsin or subjected to mechanical lysis, at 9 days post-infection. Graph shows parasite kinetics of growth in HFF for pepsin-activated versus inactivated bradyzoites. (C) Representative fluorescent images of replication in HFF, Caco-2, and HIEC-6 infected with activated bradyzoites at 7 days post-infection. Graph shows parasite kinetics of growth in the three different cell lines. Schematic representation in (A) was created with BioRender.com.

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