|Millicell Organotypic Cell Culture Procedure|
|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
|Effects of cimetidine on the biological behaviors of human gastric cancer cells|
Jiang CG, Liu FR, Xu HM, Wu T, Gao J.
Zhonghua Yi Xue Za Zhi. 2006 Jul 11;86(26):1813-6 2006
|Overexpression of TIMP-1 mediated by recombinant adenovirus in hepatocellular carcinoma cells inhibits proliferation and invasion in vitro|
Xia D, Yan LN, Xie JG, Tong Y, Yan ML, Wang XP, Zhang MM, Zhao LY.
Hepatobiliary Pancreat Dis Int. 2006 Aug;5(3):409-15. 2006
|Glucose utilization by the retinal pigment epithelium: evidence for rapid uptake and storage in glycogen, followed by glycogen utilization.|
Senanayake P, Calabro A, Hu JG, Bonilha VL, Darr A, Bok D, Hollyfield JG.
Exp Eye Res. 2006 Aug;83(2):235-46. Epub 2006 May 11. 2006
|Development of mouse embryos co-cultured with polarized or non-polarized uterine epithelial cells using sequential culture media.|
Azadbakht M, Valojerdi MR, Mowla SJ.
Anim Reprod Sci. 2006 Jul 27 2006
|A comparison of polarized and non-polarized human endometrial monolayer culture systems on murine embryo development.|
Baghaban Eslami Nejad MR, Rezazadeh Valojerdi M, Kazemi Ashtiani S.
J Exp Clin Assist Reprod. 2005 Apr 19;2(1):7. 2005
|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
|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
|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
|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
|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 interested in doing Organotypic Cell Culture. Does Millipore have a product that I can use for this?||Yes. The Millicell Cell Culture Plate Inserts are used widely in this application...namely the Millicell CM. This device contains a Biopore ( hydrophilized teflon) membrane that is optically transparent. Explants attach readily to the surface of the insert and our new low profile version allows for easy manipulation of the sample for a wide range of cytological and electrophysiological evaluations. The standard Millicell CMs are catalogue numbers PICM01250 and PICM03050. The new low rise version is available under the catalogue number PICMORG50.|
|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.|
|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.|
|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
|What are typical TEER readings for Caco-2 monolayers?||Typical in-house TEER readings for a 21 day Caco-2 cell monolayer is @ 300-500 ohms*cm2.|
|Millicell 24 and Millicell 96|
|Millicell Culture Plate Inserts|
|Millicell-CM Low Height Culture Plate Inserts User Guide|
|Millicell-ERS User Guide|