Cultrex Reduced Growth Factor Basement Membrane Extract, Type R1
Cultrex Reduced Growth Factor Basement Membrane Extract, Type R1 Summary
• Qualified for use in organoid cell culture
• Improves take-rate and growth in difficult organoid systems
• Rich in Entactin
• Quality controlled for peformance consistency
Why Use Cultrex RGF BME, Type R1?
Cultrex Basement Membrane Extract (BME) is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm (EHS) tumor. This extract provides a natural extracellular matrix hydrogel that polymerizes at 37°C to form a reconstituted basement membrane. Cultrex BME Type R1 provides a proprietary formulation that has higher tensile strength when compared to our other products Cultrex BME, Cultrex BME Type 2 and Cultrex BME Type 3. Our new Cultrex BME Type R1 has a higher concentration of entactin, one of the BME components that connects laminins and collagens reinforcing the hydrogel structure. Cultrex BME Type R1 has been specifically designed to culture tissue organoids.
Basement membranes are continuous sheets of specialized extracellular matrix that form an interface between endothelial, epithelial, muscle, or neuronal cells and their adjacent stroma and that play an essential role in tissue organization by influencing cell adhesion, migration, proliferation, and differentiation. The major components of BME include laminin, collagen IV, entactin, and heparin sulfate proteoglycans.
Protocols Utilizing Cultrex RGF BME, Type R1 for Organoid Cell Culture.
Organoid-qualified Cultrex RGF BME matrices are ideal scaffolds for organoid and 3D cell culture. R&D Systems offers multiple varieties for organoid culture with Cultrex RGF BME, Type R1 being recommended for more difficult to establish and grow tissue types. Listed below are protocols, designed by our research and development groups, for organoid culture that feature our organoid qualified Cultrex RGF BME matrices.
Protocol for Mouse Enteric Organoid Culture.
Protocol for Human Intestinal Organoid Culture.
Protocol for Human Liver Organoid Culture.
Protocol for Human Lung Organoid Culture.
Protocol for Harvesting Organoids for Biochemical Analysis.
Gelling Assay - Cultrex RGF BME, Type R1 gels in less than 30 minutes at 37°C, and maintains the gelled form in culture medium for a minimum of 14 days at 37°C.
Dome Assay Cultrex RGF BME, Type R1 forms and maintains stable 3-D dome structures on cell culture plates.
Tube Formation Assay - Cultrex RGF BME, Type R1 supports formation of capillary-like structures by human (HBMVEC; HUVEC) or mouse (SVEC4-10) endothelial cells.
For research use only. Not for diagnostic use.
What is the Tube Formation Assay?
The Tube Formation Assay is based on the ability of endothelial cells to form three-dimensional capillary-like tubular structures when cultured on a hydrogel of reconstituted basement membrane, such as Cultrex Basement Membrane Extract (BME).
What are the advantages of the Tube Formation Assay?
The Tube Formation Assay is the most widely used in vitro angiogenesis assay. The assay is rapid, inexpensive and quantifiable. It can be used to identify potentially angiogenic and anti-angiogenic factors, to determine endothelial cell phenotype, and to study pathways and mechanisms involved in angiogenesis. It can be performed in a high throughput mode to screen for a large number of compounds.
What cell types can be used in the Tube Formation Assay?
The Tube Formation Assay is specific for endothelial cells, either primary cells or immortalized cell lines. Only endothelial cells form capillary-like structures with a lumen inside. Other endothelial cell types form other structures.
What are the variables associated with the Tube Formation Assay?
The major variables associated with tube formation are composition of the Cultrex Basement Membrane Extract (BME) hydrogel, thickness of the hydrogel, cell density, composition of angiogenic factors in the assay medium, and assay period.
Which Cultrex Basement Membrane Extract (BME) should I use for the Tube Formation Assay?
Cultrex Reduced Growth Factor BME (RGF BME) is generally used for testing compounds that promote angiogenesis because formation of capillary-like structures (tubes) is significantly less compared to non-growth factor reduced varieties of Cultrex BME. The Cultrex In Vitro Angiogeneis Assay (Tube Formation) includes a qualified production lot of Cultrex RGF BME that exhibits reduced background tube formation in the absence of angiogenic factors.
How do I reduce spontaneous formation of tubular structures on Cultrex BME in the absence of angiogenic factors?
Primary endothelial cells, such as Human Umbilical Vein Endothelial Cells (HUVECs) form capillary-like structures in the absence of added angiogenic factors less often than immortalized endothelial cells. Generally, reducing the number of cells per cm2 plated onto Cultrex BME will result in less background or spontaneous tube formation. Titrate the number of cells and find optimal conditions for your specific cell line. When endothelial cells fully form capillary structures in response to angiogenic activators, but not in their absence, you may proceed.
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