Matrigel

Kepone (chlordecone) and potent estrogens in primary cultures of adult rat hepatocytes on Matrigel. Toxicol Lett 71 183-196. 

Novikova LN, Mosahebi A, Wiberg M, Teienghi G, Kellerth JO, Novikov LN (2006) Alginate hydrogel and matrigel as potential cell carriers for neurotransplantation. Journal of Biomedical Materials Research Part A 77A 242-252. 

Load 150(jlL of cells from each sample onto the upper well or inserts containing the Matrigel, and 0.6 mL medium containing chemokines in the presence or absence of inhibitors or DMSO in the bottom well. 

Fig. 21. In vivo Ti weighted spin echo MR image (7.05 T) of EPCs labeled with [GdHPD03A(H20)] (on the left). The cells are dispersed into a subcutaneous matrigel plug seven days after the implantation. On the right is shown the control image of the same in the absence of the paramagnetic label.

For non-suspension cultures, suitable matrices include liquid overlay on agarose (59), Matrigel , or Cultrex (48, 92). More recently, micropatterned arrays have been developed for adherent 3-D spheroid cultures (56) and have been used to show reduced chemosensitivity of colorectal carcinoma cells to irinotecan (58). In some cases, 3-D cultures can be enhanced by the addition of host cells. This increases complexity, but inevitably decreases flexibility and speed of analysis. However, important insights into the role of host cells have emerged stromal cells modify the gene expression and response of many tumor cell types to chemotherapeutic agents (93) and tumor-associated myofibroblasts can enhance tumor invasiveness (94). 

Rotating culture vessels such as simulated microgravity systems are primarily used to study 3-D tumor growth and differentiation. However, mixed cell populations combined with matrix proteins can be used to generate a complex microenvironment in which cell-cell interactions and invasion can be measured (95). A similar system has also been described for the coculture of endothelial cells, myofibroblasts, and tumor cell clusters embedded in Matrigel . Differential labeling of the cell populations enables their invasion and the effects of inhibitors to be measured (96). 

We and others have based invasion assays on preformed tumor spheroids generated either in hanging drops (88) or other nonadherent systems (61). These are then transferred to matrix monolayers or embedded in Matrigel and can be analyzed qualitatively or quantitatively, usually by microscopy and image analysis (see also Chapter 16). A96- or 1,536-well spheroid-based... 

D invasion from spherical structure into Matrigel 3-D from 3-D Reproducible, semiautomated, single spher-oids/well, ECM components can be varied, HT in 96-well format Central positioning of spheroids essential for automated quantification, Matrigel needs to be added slowly to maintain spheroids central location (61,78, 130, 131)... 

Dilute Matrigel to 125 pg/mL (see Note 5) in serum-ftee medium using precooled pipette tips. 

Matrigel total protein concentration is approximately 10 mg/ mL, but the precise concentration is specified on the data sheet that accompanies each batch. 

Matrigel (growth factor reduced), Becton Dickinson. 

Matrigel stock and ready-to-use solution. Thaw a 10-mL vial of Matrigel overnight at 4°C. Aliquot into sterile tubes (0.5 mL/aliquot) using refrigerated plastic pipettes and store at -20°C. Dissolve a 0.5 mL aliquot into 150 mL DMEM to obtain the ready-to-use solution. 

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

Transwell-Medlated Matrigel to Assess NSC Migration/Invasion Potential... 

Cell migration and invasion are cardinal properties of NSCs and CSCs, respectively. Assessing the effect of drugs on these functional parameters is therefore of paramount preclinical relevance. A Matrigel invasion assay, performed in Transwell chambers, represents the most advantageous and efficient method. 

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

Seed cells at a density of 2x lO cells/insert onto the layer of Matrigel in proliferation medium with or without the compound under testing for the appropriate time. 

Fig. 2. Matrigel plug assay results. Matrigel was implanted subcutaneously into athymic mice on d 0 (8-10 mice per group). Daily BMS-275291 oral treatments began on d 0 and continued to d 6. Matrigel plugs were harvested on d 7 and processed for histochemical analyses. Results are expressed as % Inhibition SE (% Relative to Untreated Control).

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