|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
|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
|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
|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
|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
|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
|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.
|Millicell Culture Plate Inserts|
|Millicell Hanging Cell Culture Inserts|
|Millicell-CM Low Height Culture Plate Inserts User Guide|
|Millicell-ERS User Guide|
|Millicell® Hanging Cell Culture Inserts|