Post on 02-Dec-2021
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Phase-Guide™
Caco-2 tubule
MediumCollagen gel
Well layout
To perfuse your cultures: place on the OrganoFlow® for programmable rocking.
Please check protocols and the rocker manual on our website for instructions.
ECM Channel Perfusion Channel Observation Window
Gel Inlet/Outlet (green)Used to access the collagen gel.
Top Perfusion Channel inlet and outlet (red), bottom Perfusion Channel inlet and outlet (orange) Inlets used to add medium, reagents or compounds to the Caco-2 tubules. (Apical or basal side)
The Observation Window (blue)Used for imaging your culture.This is where the three channelscome together and makecontact.
Visit mimetas.com/support for the latest protocols.
OrganoPlate® Caco-2 in a nutshell Assay Ready Gut Tubules
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time (days)
TEER
(ohm
*cm
2)
800
600
400
200
0
Caco-2 over time
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20 hours exposure of healthy Caco-2 tubules to Staurosporine dilutions in the OrganoPlate®
Nor
mal
ized
TEE
R
0:02 0:05 0:11 0:22 0:45 1:30 3:00 6:00 12:00 0:00
1.2
1
0.8
0.6
0.4
0.2
0
0uM9,7nM39nM156nM625nM2,5uM10uMnegative
Experiments in the OrganoPlate® Caco-2
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Culture readout examples
The four steps to get started with the OrganoPlate® Caco-2
Applications
CellsECM-gelNumber of tissue chipsNumber of control chipsCompound access to tissue tubulesMicro!uidic lane widths Internal volumesGel-medium interface surfacePlate format
Materials
Micro!uidic lane height PhaseGuide™ dimensions Medium volumePerfusion
Compatible assays
Readouts
Drug-induced toxicity, Intestinal Permeability, DDI studies with P-gp transporter Caco-2 cell line, Human Caucasian colon adenocarcinoma, Wild-TypeCollagen 1 (Rat Tail)382Apical & basal (in- and outside)
Top and bottom perfusion lane: 320 μm. Middle lane: 360 μmTop and bottom perfusion lane: 1.1 μL. Middle lane: 0.87 μL0.5 mm2
SBS-standard 384-well plate, 127.76 x 85.48 x 14.8 mm, (l x w x h) (17.3 mm height with lid)Top plate: virgin polystyrene. Plate bottom: optical quality 170 μm glass (1.5H coverslip thickness). Micro"uidics: glass, proprietary polymers, biocompatible and low compound-absorbing.220 μm100 x 55 μm (w x h)15-75 μL in each wellGravity driven, pump free, typical shear forces in tubule 0 - 3 dyne/cm2
• Immunostaining • DNA isolation • RNA isolation• Cell viability assays• Transport assays
Imaging (phase contrast, wide#eld "uorescence, confocal), plate reader (absorption, "uorescence, luminescence), sampling (ELISA, PCR, sequencing, MS, biochemistry)
• Migration assays• Barrier integrity assay• Western blot• Mass spectometry• Adhesion assays
Product speci"cations
• ELISA• TEER
Unpack the OrganoPlate® Caco -2 and remove shipment medium accordingto protocol
Incubate and perfuse the OrganoPlate® Caco-2 overnight
Perform your experiment
Check for the latestprotocols:
mimetas.com/support
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