3d optec models  (OPTEC Inc)

Journal: Scientific Reports

Article Title: 3D proximal tubule-on-chip model derived from kidney organoids with improved drug uptake

doi: 10.1038/s41598-022-19293-3

Figure Lengend Snippet: Proximal tubule epithelial cells in kidney organoids. ( a ) Schematic overview of process used to create 3D organoid-derived proximal tubule epithelial cell (3D OPTECs)-on-chip models. ( b ) Confocal image of kidney organoid (tissue-cleared) stained for PODXL + (red), LTL + (green), and CDH1 + (magenta), scale bar = 100 μm. ( c ) Schematic showing marker localization in different segments of the nephron. ( d ) Higher magnification, confocal image of kidney organoid (cryo-sectioned) stained for LTL + and CDH1 + regions, scale bar = 10 μm. ( e – g ) Confocal images of kidney organoid (cryo-sectioned) stained for AQP1 + , LRP2 + , and SLC3A1 + (magenta) in nephron segments, scale bars = 50 μm. ( h ) Heat map comparing transporter expression of OPTECs isolated from kidney organoids at days 21, 35, 49, 84, and 105. ( i , j ) Confocal images of kidney organoid (day 49, cryos-ectioned) stained for OCT2 + (magenta), LTL + (green), and CDH1 + (cyan), scale bar = 100 μm. ( k , l ) Higher magnification, confocal images of kidney organoid (day 49, cryo-sectioned) stained for OCT2 + (magenta) and LTL+ (green), scale bar = 10 μm.

Article Snippet: The increased transporter expression and polarization observed in our 3D OPTEC models after 14 days of perfusion directly translated to an observed increase in drug uptake and normalized LDH release by these tubules.

Techniques: Derivative Assay, Staining, Marker, Expressing, Isolation

Journal: Scientific Reports

Article Title: 3D proximal tubule-on-chip model derived from kidney organoids with improved drug uptake

doi: 10.1038/s41598-022-19293-3

Figure Lengend Snippet: 3D OPTEC-on-chip model. ( a ) Schematic views showing the processing steps used to create multiplexed, 3D OPTEC-on-chip models. ( b ) Corresponding images (left to right) of a representative chip after placing the channel templates, infilling the chip with ECM, removing the templates to create two-colocalized channels, and seeding one channel with OPTECs to create a 3D proximal tubule. ( c ) Confocal image of 3D OPTEC tubule showing actin (red) and DAPI (blue), scale bar = 50 μm. ( d ) Cross-sectional image of 3D OPTEC tubule (day 14, perfusion) highlighting the formation of a confluent monolayer, scale bar = 50 μm. ( e ) Brightfield image of 3D OPTEC tubule upon reaching confluency (day 7), scale bar = 75 μm. ( f ) OPTEC tubule (day 14, perfusion) stained for Na + /K + ATPase (green), LTL (magenta), and DAPI (blue). ( g ) OPTEC tubules exhibit proper apical polarization of primary cilia marker, acetylated alpha tubulin (red). ( h , i ) Basement membrane proteins laminin (red) and Col IV (green) are deposited by OPTECs on chip. ( j ) Proper expression of AQP1 (yellow) is observed in OPTEC tubules. ( f , j ) scale bars = 20 μm. ( k ) SEM image highlighting primary cilia on OPTEC, scale bar = 5 μm. ( l ) SEM image of OPTEC brush border, scale bar = 20 μm. ( m ) TEM image of brush border, scale bar = 1 μm.

Article Snippet: The increased transporter expression and polarization observed in our 3D OPTEC models after 14 days of perfusion directly translated to an observed increase in drug uptake and normalized LDH release by these tubules.

Techniques: Staining, Marker, Membrane, Expressing

Journal: Scientific Reports

Article Title: 3D proximal tubule-on-chip model derived from kidney organoids with improved drug uptake

doi: 10.1038/s41598-022-19293-3

Figure Lengend Snippet: Improved transporter expression and polarization of 3D OPTECs-on-chip. ( a ) Heat map showing OCT2, OAT1, and OAT3 transporter expression for OPTECs and PTEC-TERT1s-on-chip (day 0, as seeded), after achieving confluency (day 7, perfusion), and one-week after achieving confluency on chip (day 14, perfusion). ( b ) General transporter analysis comparing OPTEC tubules normalized by PTEC-TERT1 tubules after day 14 of perfusion on chip, one sample t test, n = 6 tubules across 3 batches of OPTECs, *p < 0.05, **p < 0.01, ***p < 0.005. ( c , d) Immunofluorescence images showing OCT2 (green) and DAPI (blue) staining in OPTEC tubules after day 14 of perfusion on chip. ( e–f ) Immunofluorescence images of showing localization of OAT3 (green) and DAPI (blue) in OPTEC tubules after day 14 of perfusion on chip ( g , h ), Immunofluorescence images showing OCT2 (green) and DAPI (blue) staining in PTEC-TERT1 tubules after day 14 of perfusion on chip, and ( i , j ) Immunofluorescence images of showing localization of OAT3 (green) and DAPI (blue) in PTEC-TERT1 tubules after day 14 of perfusion on chip, scale bars = 20 μm.

Article Snippet: The increased transporter expression and polarization observed in our 3D OPTEC models after 14 days of perfusion directly translated to an observed increase in drug uptake and normalized LDH release by these tubules.

Techniques: Expressing, Immunofluorescence, Staining