Millicell® Cell Culture Inserts

Optimized cell growth, attachment and differentiation

Overview

Specifications

Ordering Information

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Millicell Standing Inserts Clear Sorting & Filtering
Catalog Numbericon Mediaicon Chemistryicon Pore Size (µm)icon Filtration Area (cm²)icon Configurationicon Qty/Pkicon
PI8P01250IsoporePolycarbonate8.00.624-well plate50 Show Pricing & Availability
PICM01250BioporeHydrophilic PTFE0.40.624-well plate50 Show Pricing & Availability
PICM03050BioporeHydrophilic PTFE0.44.26-well plate50 Show Pricing & Availability
PICM0RG50BioporeHydrophilic PTFE0.44.26-well plate50 Show Pricing & Availability
PIHA01250MF-MilliporeMixed Cellulose Esters0.450.624-well plate50 Show Pricing & Availability
PIHA03050MF-MilliporeMixed Cellulose Esters0.454.26-well plate50 Show Pricing & Availability
PIHP01250IsoporePolycarbonate0.40.624-well plate50 Show Pricing & Availability
PIHP03050IsoporePolycarbonate0.44.26-well plate50 Show Pricing & Availability
PITP01250IsoporePolycarbonate3.00.624-well plate50 Show Pricing & Availability
PIXP01250IsoporePolycarbonate12.00.624-well plate50 Show Pricing & Availability

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Millicell Hanging Inserts preloaded in receivers Clear Sorting & Filtering
Catalog Numbericon Chemistryicon Pore Size (µm)icon Filtration Area (cm²)icon Configurationicon Qty/Pkicon
PIRP12L04Polyethylene Terephthalate1.00.3324-well plate48 Show Pricing & Availability
PISP12L04Polyethylene Terephthalate3.00.3324-well plate48 Show Pricing & Availability

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Millicell Hanging Inserts Clear Sorting & Filtering
Catalog Numbericon Chemistryicon Pore Size (µm)icon Filtration Area (cm²)icon Configurationicon Qty/Pkicon
PIEP12R48Polyethylene Terephthalate8.00.3324-well plate48 Show Pricing & Availability
PIEP15R48Polyethylene Terephthalate8.01.112-well plate48 Show Pricing & Availability
PIEP30R48Polyethylene Terephthalate8.04.56-well plate48 Show Pricing & Availability
PIHT12R48Polyethylene Terephthalate0.40.3324-well plate48 Show Pricing & Availability
PIHT15R48Polyethylene Terephthalate0.41.112-well plate48 Show Pricing & Availability
PIHT30R48Polyethylene Terephthalate0.44.56-well plate48 Show Pricing & Availability
PIMP12R48Polyethylene Terephthalate5.00.3324-well plate48 Show Pricing & Availability
PIMP15R48Polyethylene Terephthalate5.01.112-well plate48 Show Pricing & Availability
PIMP30R48Polyethylene Terephthalate5.04.56-well plate48 Show Pricing & Availability
PIRP12R48Polyethylene Terephthalate1.00.3324-well plate48 Show Pricing & Availability
PIRP15R48Polyethylene Terephthalate1.01.112-well plate48 Show Pricing & Availability
PIRP30R48Polyethylene Terephthalate1.04.56-well plate48 Show Pricing & Availability
PISP12R48Polyethylene Terephthalate3.00.3324-well plate48 Show Pricing & Availability
PISP15R48Polyethylene Terephthalate3.01.112-well plate48 Show Pricing & Availability
PISP30R48Polyethylene Terephthalate3.04.56-well plate48 Show Pricing & Availability

* For attachment-dependent cells, this membrane needs to be coated with an extra cellular matrix.

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Tissue Culture Treated Cell Culture Receiver Plates Clear Sorting & Filtering
Catalog Numbericon Device Materialicon Number of Wellsicon Filtration Area (cm²)icon Recommended Media Volume (mL)icon Qty/Pkicon Packagingicon
PIMWS0650Polystyrene69.51.9–4.350Individually blister packaged Show Pricing & Availability
PIMWS1250Polystyrene123.80.9–1.850Individually blister packaged Show Pricing & Availability
PIMWS2450Polystyrene241.90.6–1.650Individually blister packaged Show Pricing & Availability

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Accessories Clear Sorting & Filtering
Catalog Numbericon Descriptionicon Qty/Pkicon
MERS00002Millicell ERS-2 Voltohmmeter1 Show Pricing & Availability
MERSSTX01Replacement Electrodes, 1 pair1 Show Pricing & Availability

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    Documentation

    Brochure

    Title
    Advanced cell culture systems for every challenge.
    Multiwell solutions for discovery research and sample prep

    Technical Info and Required Licenses

    Title
    Millicell Organotypic Cell Culture Procedure

    Data Sheet

    Title
    Millicell Cell Culture Plate Inserts: For attachment-dependent, suspension cell or organotypic growth with apical and basolateral access
    Millicell Inserts and Plates

    References | 55 Available | See All References

    Reference overviewApplication
    Propagation of human embryonic stem cells in a microporous membrane-based indirect co-culture system
    Kelsey Albert, Steven Sheridan, Louise Laurent, Igor Ulitsky, Ron Shamir, Jeanne Loring, & Raj R. Rao
    Biochemical and Biophysical Research Communications 2010

    Astrocyte growth effects of vascular endothelial growth factor (VEGF) application to perinatal neocortical explants: Receptor mediation and signal transduction pathways
    Nina Mani, Alfia Khaibullina, Janette Krum and Jeffrey Rosenstein
    Experimental Neurology 192 (2005); 394-406 2005

    Cell Culture
    Subcellular localisation of recombinant a and g-synuclein
    Christian Specht, Cezar Tigaret, George Rast, Agnea Thalhammer, York Rudhard and Ralf Schoepfer
    Mol. Cell. Neurosci., 28 (2005); 326-334 2005

    Cell Culture
    Establishment of the organotypic model of amyotrophic lateral sclerosis from the SD rats' spinal cord
    Diao ZY, et. al,Beijing Da Xue Xue Bao. 2005 Apr 18;37(2):134-8. Chinese.
    Beijing Da Xue Xue Bao. 2005 Apr 18;37(2):134-8. Chinese. 2005

    Cell Culture
    Neural stem cells protect against glutamate-induced excitotoxicitiy and promote survival of injured motor neurons through the secretion of neurotrophic factors
    Jeronia Llado, Christine Haenggeli, Nicholas Maragakis, Evan Snyder and Jeffrey Rothstein
    Mol. Cell. Neurosci. , 27 (2004); 322-331 2004

    Cell Culture
    Development of an in vitro blood-brain barrier model-cytotoxicity of mercury and aluminum.
    Toimela, T et. al.,Toxicol Appl Pharmacol. 2004 Feb 15;195(1):73-82.
    Toxicol Appl Pharmacol. 2004 Feb 15;195(1):73-82. 2004

    Morphological differentiation of bone marrow stromal cells into neuron-like cells after co-culture with hippocampal slice
    Abouelfetouh, Ayman, et al
    Brain Research (2004), Volume 1029, Issue 1 pp 114-119 2004

    Cell Culture
    Effect of sodium bicarbonate on extracellular pH, matrix accumulation, and morphology of cultured articular chondrocytes.
    Waldman SD, Couto DC, Omelon SJ, Kandel RA
    Tissue Engineering (2004), Nov-Dec;10(11-12):1633-40 2004

    FGF-10 plays an essential role in the growth of the fetal prostate.
    Annemarie A. Donjacour, Axel A. Thomson and Gerald R. Cunha
    Developmental Biology 261 (1): 39-54 2003

    Cell Culture
    Changes in lymphokine receptor expression and fatty acid composition of phospholipids and triacylglycerols in rat adipocytes associated with lymph nodes following a transient immune challenge.
    J. D. Priddle, C. A. Mattacks, D. A. Sadler, H. A. MacQueen and C. M. Pond
    Cell Biology International 27 (1): 23-29 2003

    Cell Differentiation

    FAQ

    QuestionAnswer
    Are the Millicell PCF inserts tissue culture treated on both sides of the membrane?Yes, they are tissue culture treated on both sides of the membrane. This allows for attachment- dependent cell culture on both sides of the membrane.
    Are the Millicell units sterile and, if so, how are they sterilized?The Millicell units are sold sterile in individual blister packs. The HA and CM units are ETO sterilized and the PC and PCF are Gamma Irradiated.
    Can cells grown on Isopore (polycarbonate) membrane be viewed under a microscope?Yes, we can observe the cells seeding on the membrane of pore size 1 um.
    Can I make membrane potential measurements with the Millicell-ERS?Yes, membrane potential measurements can be made with the ERS. Short circuit potential, however, cannot be done.
    How can I be sure that I have a confluent monolayer in my Millicell ?There are several answers to this question based on the type of membrane you are using and the equipment you have. It is possible to view, via phase contrast microscopy, actual cell layers if you are using Millicell HA, PC and PCF.

    With Millicell CM you can use bright field microscopy as well to check for confluency. Also, one way of assuring confluency is to seed your cells at optimal levels. Most cell lines do best at a density of 5x10^5 to 1x10^6 cells/ cm2. So long as you are working with a cell type (i.e. epithelial cells) that exhibits resistance due to membrane potential you can also use the Millicell ERS to confirm confluency.
    How long do the electrodes last?The electrodes will last at least 6 months with normal use and care, barring any physical damage. To prolong life of the electrodes, clean after every use and store dry.
    How should I store electrodes when not in use?When storing electrodes for long periods of time, wash the electrodes with Milli-Q water or equivalent to remove salts and proteins, then store dry. For short term storage electrodes can be stored in a buffered solution. Make sure the electrode cable plug is connected to the electrode port on the Millicell ERS meter so that the system is internally short-circuited and electrode symmetry is maintained.
    I am coating the Millicell CM with Rat Tail collagen according to your instructions and sometimes I am seeing an empty ring at the periphery of the insert where the collagen has not filled in and an empty "hole" at the center where there is no coating either. How can I avoid this from happening in the future?If you are using Type 1 collagen, it should be filter sterilized in non-denaturing alcohol prior to use in the Millicell. Also, try looking at the insert while you are coating it and tap/rotate it to enhance uniform liquid coating. The entire membrane should go clear which indicates uniform wetting/coverage of the Millicell. Also, be sure to use at least minimum volumes of 50ul for the 12mm and 350 ul per 30 mm.
    I am concerned about the possibility of introducing surfactants in cell cultures using the Millicell PCF product. Is the Polycarbonate membrane in Millicell-PCF PPVP (surfactant) free?Yes, the Polycarbonate membrane in Millicell-PCF is PPVP (surfactant) free.
    I am culturing tissue slices on the Millicell Polycarbonate unit (PIHP01250) and it appears that the media is not being retained by the unit. What is going on?The 0.4 um PCF polycarbonate membrane is tissue culture treated and, therefore, hydrophilic. In long-term culture ( >than a few to 24 hours) they will NOT retain fluid since it will slowly pass through the pores. A monolayer of epithelial cells on the membrane serves as a secondary barrier which will selectively prevent/or allow molecules to pass.

    If you are referring to an overnight culture with no media outside the Millicell this result is to be expected. If there is equal fluid heights both inside and outside the Millicell ( ~ 400 ul inside and 600 ul outside) then the fluid will be retained. Since they are doing organotypic culture you could easily adjust the inside and outside volumes to the same desire height to make sure the culture remains bathed in media.
    I am testing my Millicell ERS and cannot get a value when I take a reading. What am I doing wrong ?When testing the unit, it is very important to hold down the test button until the reading is taken, otherwise you will not get a real value. If you only push the button down and release no reading will be obtained.
    I have seeded cells on the Millicell and they are detaching from the membrane, what do I do to correct this?Make sure that the medium on the outside of the insert is lower than the level of medium on the inside of the insert. Also, if it is important to remember that certain cells lines, i.e. attachment dependent cells, often require pretreatment of the Millicell membrane in order to enhance attachment. This is usually accomplished through ECM coating...especially in the case of Millicell CM where coating is always required. Too much feeding of unconditioned media can also cause detachment in some cell types, e.g. caco-2.
    Is it necessary to treat the Millicell CM with an ECM coating if I am doing organotypic culture ?In general, it is not required that you treat the Millicell CM with an ECM coating when doing organotypic culture but this is dependent on the type of tissue being cultured and the procedure being followed. In Tech Note TN062 , we outline a procedure for organotypic culture that uses a pretreatment of the Millicell CM with poly-ornithine ( an attachment factor). Other references cited in this paper do not use this precoating procedure. When evaluating whether or not to use an attachment factor in your experiment, it is best to use as many resources as possible and decide based upon the specific experimental parameters in your situation.
    Is there a protocol to cut out the membranes from Millicell inserts?The membrane can be removed from the insert insert. This is most easy accomplished using a Diamond knife over a piece of cork.
    What are the dimensions of the Millicell units?For the 12 mm units, the height is 10.5 mm. The diameter (0D) of the base is 13mm & the top is 12 mm. The ID is 10 mm. The membrane diameter is 12mm and effective membrane area is 0.6 cm2. For the 30 mm unit, the dimensions are 13 mm height. 32mmOD base, 30 mmOD top and 27mmID. The membrane diameter is 30 mm and the effective area is 4.2 cm2.
    What cell types can be used with the Millicell ERS?The MERS will only give meaningful readings on a cell line that is know to form tight junctions e.g. epithelial cells. We strongly recommend using the ERS for those cell lines where published values exist for the expected resistance. Many cell types do not have a large enough resistance to use this technique.
    What is an ECM and why should it be used?ECM is extracellular matrix material and is used to coat certain Millicells ( e.g. Millicell CM) to enhance cellular attachment. Normally, attachment dependent cells cannot readily bind to the Biopore membrane so coating the Millicell with collagen or other ECM material will render the filter surface suitable for cell attachment. Please note: the Millicell PCF is tissue culture treated and should not require additional coating. In general, the Millicell HA does not require coating with an ECM and will support direct cell attachment.
    Why do the electrodes need to be replaced every six months?In general, electrodes have a 6 month lifetime if they are used continuously. The electrodes have a silver-silver chloride pellete that is depleted everytime it is used. It is the silver silver chloride pellete that creates the potential.
    What voltage value can I expect for my cell type?We DO NOT recommend using this function on the instrument. The Millicell-ERS will make voltage readings on only a few kinds of high-voltage generating cells (for example, it will not work on MDCK cells). We do have information on epithelial cells which typically have voltage values of 0-30mV.
    What resistance value can I expect for my cell types?Some resistance values have been published for various cell types. Resistance measurements will also depend on the type of cell culture (i.e. cell line, tissue or native cells). In general, the resistance range for epithelial cells is 10-20,000 ohms/cm2. An example of an established epithelial cell line with well defined electrical resistance is MDCK cells. MDCK cells displaying tight junctions show resistance of 5000 ohms/cm2.
    What is the porosity of the Millicell Culture Plate Inserts?The porosity of the Millicell Insert membranes are as follows: HATF - approx 80% CM - approx 80% PC and PCF - approx 10%
    What is the length of the legs on the Millicell Insert unit?The legs on the Millicell unit are approximately 1-2mm.
    What is the thickness of the Millicell-PC and -PCF membranes?The thickness of the PC and PCF membranes are 10 um.
    What is the thickness of the Millicell-CM membrane?The thickness of the Millicell-CM memnbrane is approximately 50 um.
    What is the overall height of the Millicell-CM Organotypic insert?The overall wall height including the feet is 5 mm.
    What Volume of media should be used on the inside and outside of the 12 mm and 30mm Millicell cell culture insert devices?Inside and outside liquid heights are critical and must be adjusted until equal. The suggested volumes below can vary due to plate well volume variations allowed by different cell culture plate manufacturers.

    12 mm inserts:
    Inside: 0.4 mL
    Outside: 0.6 mL
    Which ECM coating do we recommend for endothelial cells?We recommend the Type 1 Rat Tail Collagen for most endothelial cells. It is recommended to be done with ethanol wetting and is a very cost efficient way to do ECM and seems to be well tolerated by most endothelial cells.
    What is the difference between Millicell PC and PCF?Both Millicells have polycarbonate membranes and are frequently used in chemotaxis and transport studies. PC's are recommended ONLY for suspension cell culture. PCF's are tissue culture treated and therefore recommended for attachment dependent cell culture.
    What is the advantage of using Millicell for cell culture applications ?Studies have shown that cells actually grow better on microporous membranes than on the artificially smooth surfaces of standard cell culture plasticware. We have found that cells grown on Millicell Cell Culture Plate Inserts produce higher cell densities and greater differentiation than cells grown in standard setups. Cells can be constantly bathed and fed on both their apical and basolateral surfaces and ions and molecules can pass freely across the membrane. Millicells also offer the researcher a more humane approach to animal testing since cell culturing with the units can offer alternatives to live animal studies for many toxicological studies.
    What size tissue culture plate do I use with the Millicell devices?The 12mm Millicell insert can be used with a standard 24 well plate, and the 30mm insert is used with a 6 well plate.
    What is the height of the standard Millicell feet ? ( i.e. how high does the Millicell sit off the floor of the plate?)The Millicell feet are 1-1.5 mm high.

    User Guides

    Title
    Millicell 24 and Millicell 96
    Millicell Culture Plate Inserts
    Millicell Hanging Cell Culture Inserts
    Millicell-CM Low Height Culture Plate Inserts User Guide
    Millicell-ERS User Guide

    C10504Optimized cell growth, attachment and differentiation

    With Millicell inserts, attachment or suspension cells can access media from both their apical and basolateral sides. Cell growth, structure, and function more closely mimic what occurs in vivo. In addition, Millicell inserts make it possible to study both sides of the cell monolayer.

    Millicell inserts are available for 24-, 12-, or 6-well plates. The inserts are easily prepared for SEM and TEM visualizing techniques, and they are compatible with cellular and/or fluorescent stains.

    Insert Formats

    New
    Millicell Hanging Inserts: Preloaded in receivers or available standalone

    • For co-culturing and permeability assays
    • Unique design allows easier basolateral access than other hanging inserts with less risk of contamination
    • Available in 5 pore sizes and 3 diameters, including a 1 µm pore size that is optically transparent for better visualization by microscopy
    • NEW: Available preloaded in 24-well receiver plates or as individual inserts in 3 diameters

    Millicell Standing Inserts

    • Promotes excellent cell growth and provides an exceptional opportunity for cell studies
    • Available with Biopore (PTFE) membrane, MF-Millipore (mixed cellulose esters) membrane, and polycarbonate membrane

    Millicell Organotypic Insert

    • For high cell viability and superior study of three dimensional explant structure
    • Lower height allows them to fit inside a standard petri dish
    • The Biopore™ (PTFE) membrane provides high viability—for as long as 40 days—and excellent trans-membrane oxygen transport
    • The membrane is optically clear and optimized for long-term organotypic explant maintenance

    Membrane Types

    Biopore Membrane (hydrophilic PTFE)

    For low protein-binding, live cell viewing, and immunofluorescent applications

    MF-Millipore™ Membrane (mixed cellulose esters)

    For exceptional anatomical and functional polarization

    Isopore™ Membrane (polycarbonate)

    For growth of attachment-dependent cells without matrix

    PET Membrane (polyethylene terephthalate)

    For growth of attachment-dependent cells without matrix

    With Millicell® inserts, adherent or suspension cells can access media from both their apical and basolateral sides. Cell growth, structure, and function more closely mimic what occurs in vivo. In addition, Millicell® inserts make it possible to study both sides of the cell monolayer.

    Features & Benefits

    • For co-culturing and permeability assays
    • Unique design allows easier basolateral access than other hanging inserts with less risk of contamination
    • Available in 5 pore sizes and 3 diameters
    • 1 µm pore size is optically transparent for better visualization by microscopy

    Applications

    Cell Attachment, Cell Growth, Cell Differentiation , Immunocytochemistry

    Product Performance

    For more information visit: www.emdmillipore.com/cellculture