Matrigel coating

Figure 12 Dose- and time-dependent induction of CYP3A activity in human cultured hepatocytes incubated with rifampicin. Cells cultured on Matrigel-coated 96-well plates were incubated with increasing doses of rifampicin and CYP3A was determined by probing the cells with DFB prior to RNA isolation. Abbreviation DFB, [3-[(3,4-difhiorobenzyl)oxy]-5,5-dimethyl-4-[4-(methylsulfonyl)phenyl]furan-2(5/F)-one].

Moroi M, Okuma M, Jung SM Platelet adhesion to collagen-coated wells analysis of this complex process and a comparison with the adhesion to matrigel-coated wells. BiochimBiophys Acta 1137 1-9, 1992... 

In most cases, hPSC cultmes on microcarriers require a Matrigel coating and/or MEF-CM. These conditions are not xeno-free culture conditions and make it difficult to use hPSCs in clinical applications. 

Split hESC/hIPSC as described in Subheading 3.1, steps 3 and 4, but now divide the colonies from one confluent well of a 6-well plate to 4 wells of a gelatin/Matrigel coated 6-well plate containing 3 mL of hPSC growth medium per well to allow maintenance of pluripotency. 

Coat the wells of 96-well plates with Matrigel overnight. 

Coat overnight with Matrigel the required number of 6-well Transwell inserts containing a polycarbonate 8.0 pm pore membrane. 

Resuspend and seed in 60 mm dishes coated with Matrigel... 

In regards to culturing hepatocytes in a 96-well plate format, we have adopted the same conditions that we used when culturing cells in 60-mm dishes and 24-well plates (12) and simply scaled them down to a 96-well plate format. The 96-well plates are precoated with Matrigel and are commercially available (Collaborative Biomedical Products, Boston, Massachusetts, U.S.) or, alternatively, normal plates can be coated with diluted Matrigel and dried overnight (27). Hepatocytes cultured on collagen-coated 96-well plates have also been reported to be suitable for CYP induction (28). 

Dilute Matrigel in cold distilled water to 50 /xg in 50-100 /xl. Apply 25-50 /xg Matrigel to the filters, dry under hood, and reconstitute with serum-free medium. Place a solution of chemoattractant in the lower compartment of the Boyden chamber (in the absence of chemoattractant, very little cell migration occurs over a 6-h period). Place the coated filters in Boyden chambers, and close the chamber. Add to the upper chamber 2 to 3 x 10 cells in appropriate medium containing 0.1% BSA. Incubate at 37°C in 5% CO2 for 4-6 h. Remove the cells from the upper surface of the filter by wiping with a cotton swab or by passing them on tissue paper. Fix the filter (methanol or ethanol) and stain (haematoxylin-eosin or toluidine blue). Count the cells from various areas of the lower filter surface. Alternatively, migrated cells can be solubilized... 

A simple variation on the method described above was used by Tullberg et al. (1989) to select B16 melanoma cells of increased invasive (and metastatic) ability. Filters with pores of 10 /.tm are employed to avoid a selection for pore penetration, and coated with a thick layer (approximately 1 fim) of Matrigel , a reconstituted BM derived from EHS tumors (see Chapter 4 for more details). Cells are plated on the coated filters at a density that is lower than that used for the migration selection, since in this case the time it takes for the cells to migrate can be considerably higher, and excessive growth on the upper side of the filter has to be prevented. Optimal conditions have to be empirically determined for each cell line. Cells that have invaded and passed to the underside of the filter are collected and expanded as described above, and further selections can be made to obtain a fairly homogenous cell population of increased invasive potential, from which clones can be derived. 

Prime/coat the channels Add diluted matrigel (1 50 in PBS, 100 pL/ well) into the oudet wells. Connect the interface and apply a shear force of 5 dyn/cm for 5 min to push PBS into the inlet weUs. Incubate the plate at 37 °C for 1 h. 

Also, the parylene-C sheath-based neural probe is coated with neurotrophic and anti-inflammatory factors loaded onto a Matrigel carrier. This encourages the ingrowth of neuronal processes for improved recording quality, reduces the immune response, and promotes an improved probe integration into the brain tissue [89],... 

Figure 6.1 Culture methods for human pluripotent stem cells (hPSCs). hPSCs can be cultured on mouse embryonic fibroblasts (MEFs) or human feeder cells. Feeder-free culturing of hPSCs is possible on xeno-containing Matrigel. Several types of feeder-free and xeno-free cultures of hPSCs can be developed on extracellular matrix (ECM)-coated surfaces and ohgopeptide-immobilLzed surfaces. hPSCs can also be cultured on polysaccharide hydrogels or on synthetic polymer surfaces by selecting a specific GAG or a specific polymer with optimal water content. Reproduced with permission from [2] Copyright 2014 Elsevier Inc.

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