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Cell-based Assays for Drug Transport

Millicell-96 cell culture insert plate is a patented 96-well device designed to support epithelial cell growth and differentiation of Caco-2 and other cell lines.

Overview

Specifications

Ordering Information

Cell-based Assays for Drug TransportClear Sorting & Filtering Show Filter
Catalog NumberDescriptionComponentsPack Size
PSHT004R1Millicell-96 Cell Culture Insert Plate
  • 96-well cell culture plate(1), Single-well feeder tray(1), 96-well receiver tray(1), lid(1)
1 Show Pricing & Availability
PSHT004S5Millicell-96 Cell Culture Insert Plate
  • 96-well cell culture plate(5), 96-well receiver tray(10), lid(5)
5 Show Pricing & Availability
PSHT004R5Millicell-96 Cell Culture Insert Plate
  • 96-well cell culture plate(5), single-well feeder tray(5), lid(5)
5 Show Pricing & Availability
PSRP004R1Millicell-96 Cell Culture Insert Plate
  • 96-well cell culture plate, single-well feeder tray, 96-well receiver tray, lid
1 Show Pricing & Availability
PSRP004R5Millicell-96 Cell Culture Insert Plate, polyethylene terephthalate, 1.0 µm
  • 96-well cell culture plate, single-well feeder tray, lid
5 Show Pricing & Availability
MACAC0RS596-well Collection Plate
  • 96-well transport tray(5), lid(5)
5 Show Pricing & Availability

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Documentation

Technical Info

Title
Cell-Based Receptor Binding Assays Performed with the MultiScreen Assay System
MultiScreen Caco-2 --Drug Transport Assembly in a 96-well system:

Data Sheet

Title
MultiScreen Assay System for High Throughput Cell-based Transport

Posters

Title
Automation of ADME Applications

References

Reference overviewApplication
Use of ADME screening to conserve resources in drug discovery and development
Weiss, A. and Joseph Machamer
American Biotechnology Laboratory, January 2004: 16-17  2004

Induction of cytokines in a human colon epithelial cell line by Shiga toxin 1 (Stx1) and Stx2 but not by non-toxic mutant Stx1 which lacks N-glycosidase activity.
Chisato Yamasaki, Yumiko Natori, Xun-Ting Zeng, Mari Ohmura, Shinji Yamasaki, Yoshifumi Takeda and Yasuhiro Natori
FEBS Letters 442 (2-3): 231-234  1999

Cell Culture

FAQ

QuestionAnswer
What is the well depth and maximum volume capacity of a MultiScreen plate?The well depth of a 96 well MultiScreen plate is 1.245 cm. The well depth for a 384 well MultiScreen plate is 1.2 cm. The maximum working volume of a 96 well plate is 300 ul. The maximum working volume for a 384 well plate is 100 ul.
Why do we recommend addition of 75 µL of medium apically (in the filter well) and 250 µL basolaterally (in the receiver/feeder plate) for the MultiScreen Caco-2 plate?These volumes were determined to be optimal for cell growth when feeding the cell culture every other day. In addition, with 75 µL apical, the medium level is at the same height as the basolateral volume of 250 µL. In this way we ensure that there is no positive pressure exerted on the monolayer of Caco-2 cells.
Do you recommend any specific handling when feeding the cells on the MultiScreen-Caco-2 plate?We recommend using an autoclavable 8-channel aspiration manifold. Sigma M2656. Drummond #3-000-093 Aspirate/feed apically using the apical notch and down the side of the well (slow speed if programmable).

If you are feeding manually, feel free to separate the plates to aspirate/feed basolaterally. Due to the fact that the MultiScreen-Caco-2 plate has recessed filter wells, the membrane will not be in direct contact with any surface. The filter plate has 4 feet it stands on.
Can I add lucifer yellow and drug compounds together in the same well in the MultiScreen Caco-2 plate?We do not recommend using lucifer yellow in the same well as your test compound if you are using radioactive drugs. Lucifer yellow may interfere with scintillation cocktail and give inaccurately high counts in the beta counter.
Why are the propranolol and testosterone values from Millipore’s laboratory determined on the MultiScreen Caco-2 plate lower than published values?We have speculated that the differences we observe are related more to the cell culture and analytical conditions we employ in our laboratory because the values we observe on the standard 24 well plates are also lower than published. Because we correlate well with the 24 well data generated at the same time in our laboratory, the discrepancy from published is not a concern.
Do you recommend a specific way of seeding the cells on a MultiScreen Caco-2 plate?We recommend pipetting cells at the apical notch and down the side of the well (slow speed if programmable). When seeding many plates, make sure to continually keep cells suspended.
Why is there a defined order for aspirating medium and addition of new medium when feeding the cells on the MultiScreen-Caco-2 plate?When culturing epithelial cells on filters, it is desirable to limit any pressure the medium may exert which may disrupt the cell contact with the filter. Therefore, the medium in the apical compartment should always be present when medium is present in the basolateral compartment. The preferred order is to aspirate basolaterally first, then apically, and add fresh medium back apically first, then basolaterally.
How much does the TEER manual electrode STX-100M cost that is recommended for use with the MultiScreen Caco-2 plate?The STX-100M probe is $1,200 and you would order this through World Precision Instruments. You will also need a Ohm meter, which is available from Millipore, MERS 000 01.
What lucifer yellow and drugs do you use on the MultiScreen Caco-2 plate? What are your test conditions?We use lucifer yellow dipotassium salt (LY) from Sigma (L0144) concentration of 100 µg/mL in HBSS). We read LY with a Victor Plate Reader (Wallac/PE). We use tritium labeled drugs spiked with unlabeled drug and use a Microbeta reader (Wallac/PE). We usually run the LY and drug transport experiments for 2 hours, room temperature, shaking at 60 rpm.
Can you use all 96 wells for the drug permeability assay in the MultiScreen Caco-2 plate?Typically we use all 96 wells for our assays. But as is the case for many cell based assays in 96 well plates, there may be more variability in the cell growth on the periphery of the plate. We confirm monolayer integrity by measuring the TEER on all wells prior to beginning a drug transport assay.