Application

ECM gel can used with epithelial cells, endothelial cells, muscle cells, nerve cells and tumor cells.

Biochem/physiol Actions

The ECM gel will undergo thermally activated polymerization when brought to 20-40°C, forming a reconstituted basement membrane. PC12 cells will show neurite formation within 2 days when they are grown on a thin layer of this ECM gel.

Caution

For long term storage, keep product at -20°C. The ECM gel may be stored at 2-8°C for up to 72 hours.

Components

ECM was prepared to a protein concentration of 8-12 mg/mL (in DMEM), containing laminin as a major component, collagen type IV, heparin sulfate proteoglycan, entactin, and other minor components. Addition of collagen type IV to the ECM gel increases polymerization.Product contains 50 mg/L Gentamicin Sulfate.

Preparation Note

This product is prepared from Engelbreth-Holm-Swarm sarcoma in mice; dialyzed against chloroform. Gel should be thawed overnight at 2-8°C before use and dispensed to a multiwall plate using a plate and pipettes that are pre-cooled to 2-8°C. The gel may also be diluted up to twofold with 2-8°C Dulbecco′s Modified Eagle′s Medium, and should be done before gel is added to the plate. The product will gel within 5 minutes at 20°C. For prolonged manipulations, work should be conducted below 10°C. Cells can be plated on top of a thin gel layer of 0.5 mm or cultured inside a 1 mm layer. If you culture cells inside, add the cells to the gel prior to plating at a recommended density of 3-4 x 10⁴ cells per mL. To dissociate cells from this gel, use protease in PBS without calcium, magnesium, or EDTA, at a concentration of 0.6-2.4 units per mL.