Matrigengel Matrix IPSC Level

Basement membranes are continuous sheets of specialized extracellular matrix that form an interface between endothelial, epithelial, muscle, or neuronal cells and their adjacent stroma. Basement membranes are degraded and regenerated during development and wound healing. They not only support cells and cell layers, but they also play an essential role in tissue organization that affects cell adhesion, migration, proliferation, and differentiation. Basement membranes provide major barriers to invasion by metastatic tumor cells.

Mogengel Matrigengel Matrix IPSC Level is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm (EHS) tumor. Mogengel Matrigengel Matrix IPSC Level at 37°C to form a reconstituted basement membrane. The major components of Mogengel Matrigengel Matrix IPSC Levelinclude laminin, collagen IV, entactin, and heparin sulfate proteoglycan.

MATERIAL QUALIFICATIONS
• Routine screening of mouse colony pathogensby mouse antibody product (MAP) tests
• Testing for bacteria, fungi and mycoplasmato ensure negative results
• Extensive PCR testing for a variety of pathogens including LDEV to ensure strict control of raw materials used in the production process
• Extraction from LDEV-free mouse tumor cells
• Gel stability testing at 37°C for 14 days
• Detection of endotoxin levels using serological methods
• Biological function verification of each lot (Organoid culture and differentiation experiments; Subcutaneous tumor formation test; Stem cell culture; Angiogenesis experiment etc.)

COATING PROCEDURES:
• Thaw Matrigengel Matrix overnight at 2-8°C. Refrigerator temperatures may vary, therefore it is recommended to keep Matrigengel on ice in a refrigerator during the thawing process. Thawed Matrigengel solidifies quickly at temperatures above 15°C; when working with Matrigengel, keep it on ice to prevent untimely gelling.
• There are many applications for Matrigengel which require different thicknesses and concentrations. A thick gel is needed for applications such as endothelial cell formation of capillary-like structures (Tube Formation Assay), the differentiation of rat aorta tissue into capillary-like structures (Aortic Ring Assay), epithelial organoid formation, or tumor organoid formation. Some applications, such as propagation of primary cells, require a thin layer coating and not a thick gel; therefore, the thin layer method should be used.

Thick Gel Method:
1. Thaw Matrigengel as stated above.
2. Mix Matrigengel by slowly pipetting solution up and down; be careful not to introduce air bubbles.
3. Pipette 200-300μL per cm2 onto the growth surface.
4. Place coated object at 37°C for 30 minutes.
5. Coated objects are ready for use.

Thin Layer Method (non-gelling):
1. Thaw Matrigengel as stated above.
2. Mix Matrigengel by slowly pipetting solution up and down; be careful not to introduce air bubbles.
3. Dilute Matrigengel to desired concentration in cold serum-free medium. A 1:100 dilution is recommended for the propagation of primary cells. Empirical determination of the optimal coating concentration for your application may be required.
4. Add a sufficient amount of solution to cover the entire growth surface area. A volume of 300μL per cm2 is recommended.
5. Incubate coated object at room temperature for one hour.
6. Aspirate coating solution and immediately plate cells. Do not allow coated surface to dry out.

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Background

It is a mature growth substrate for feeder-free hES culture. It's used in combination with specific media to culturize stem cells (e.g, hESC, iPSC). The screened stem cell Matrigengel Matrix provide the reproducibility and consistency required for trophoblast culture of human embryonic stem cells and induced pluripotent stem cells, and can also be used in vivo differentiation studies, such as teratoma formation.