Journal: eLife

Article Title: Human immunocompetent Organ-on-Chip platforms allow safety profiling of tumor-targeted T-cell bispecific antibodies

doi: 10.7554/eLife.67106

Figure Lengend Snippet: ( A ) Diagram of Colon and Duodenum-Intestine chip seeding, beginning with fragmented primary human organoids seeded into the epithelial channel of the chip. Primary intestinal endothelial cells, either colon or small-intestinal depending on corresponding epithelial tissue, are seeded into the vascular channel and the chip is cultured to maturity under flow and mechanical deformations. ( B ) Immunofluorescence staining of CEA and nuclei in colon or dudodenum chips versus matched colon or duodenum organoids. ( C ) Immunohistochemistry analysis of CEA (brown coloration) in a conventional, static model of the intestinal barrier: organoid-derived intestinal cell seeded on ECM-coated transwell membranes. ( D ) Flow cytometry-based quantification of CEA binding sites expressed by intestinal barriers cultured in transwells. High CEA-expressing cancer cell lines MKN5 and A549 serve as positive controls. ( E ) Treatment of PBMC with CEA-targeted TCBs in the absence of target does not induce activation, confirming target-dependent mode of toxicity observed in the Intestine-Chips.

Article Snippet: Human Large Intestine Microvascular Endothelial Cells (cHIMEC) (Cell Systems) were thawed at passage five and cultured in Endothelial Cell Growth Medium (EGM-2MV) (PromoCell) supplemented with Endothelial Cell Growth Medium MV2 Supplement Pack (PromoCell) and 1% v/v primocin (InvivoGen).

Techniques: Cell Culture, Immunofluorescence, Staining, Immunohistochemistry, Derivative Assay, Flow Cytometry, Binding Assay, Expressing, Activation Assay