Matrigel preparation

These problems can be circumvented by using a reconstituted BM. The most widely used material is known as Matrigel , an extract of a murine tumor which produces massive amounts of BM (Kleinman et al., 1986). In fact, MatrigeF is a mixture of BM components (mostly laminin) prepared from the murine EHS tumor of C57/black mice grown in vivo. Once extracted from the tu-... 

Matrigel is the trade name (BD Biosciences) for a solubilised basement membrane preparation extracted from the Engelbreth-Hohn-Swarm mouse sarcoma, a tumor rich in extracellular matrix proteins. This mixture resembles the complex extracellular environment found in many tissues and is used as a substrate for cell culture. 

Using a 1-mL syringe collect the contents of the Eppendorf tube with the appropriate mixture of buffer, Matrigel, and chemoattractant (such as EGF) prepared in step 3.1 and load the 33-gauge experimental needles slowly until 2-3 drops of liquid are expelled from the needle tip (rrrNote 3). 

The ECM is prepared on ice once the Matrigel is added, subsequent steps/protocols are completed swiftly to ensure the ECM does not start to polymerize before the fibroblasts/spheroids are added. 

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