What kinds of tumor cells or biopsy specimens grow in vivo with Cultrex® BME?

Many cell lines and tumor biopsy specimens (usually cut into small fragments) have been found to grow in vivo when implanted with Cultrex® BME. These include melanoma, intestinal, prostate, breast, lung, renal, and liver cancers as well as the 3T3 mouse embryonic fibroblast cell line.

How does Cultrex® Basement Membrane Extract (BME) promote cell differentiation?

All epithelial and endothelial cells are in contact with a basement membrane matrix on at least one of their surfaces. By providing them with their natural matrix in vitro as a substrate for the cells that provides biological cues, the cells can assume a more physiological morphology (i.e. correct shape) and begin expression of cell-lineage specific proteins. Two-dimensional plastic surfaces, in combination with serum-containing media, cause cells to flatten, proliferate and de-differentiate.

How should Cultrex Basement Membrane Extract (BME) be stored and handled?

For best practices for handling our BME products, please refer to our helpful guide at the following link: https://www.bio-techne.com/reagents/cell-culture-reagents/cultrex-bme-and-ecm-proteins/cell-culture-matrix-for-your-research

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.

Can cells grown in Basement Membrane Extract (BME), such as Catalog # 3433-005-01, be used in Flow Cytometry?

For preparation of a single cell suspension for applications such as FACS and re-culturing, digestion of the matrix and cell-cell bonds may be facilitated using enzymes such as Trypsin or Dispase. Our CellSperse, cat# 3450-048-05, is similar to Dispase, and can also be used for this purpose. However, some markers may be susceptible to degradation from CellSperse treatment. It is recommended to consult the literature for design of experiments for the particular cell type and markers of interest.

Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear Best Seller

Name: Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear
Brand: R&D Systems
SKU: 3433-010-01
Rating: 5 (8 reviews)

R&D Systems, part of Bio-Techne | Catalog # 3433-010-01

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Citations (50)

Product Datasheet / COA / SDS

Reduced Growth Factor Basement Membrane Extract

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Key Product Details

Features:	Cultrex Reduced Growth Factor Basement Membrane Extract (RGF BME) is an extracellular matrix hydrogel that has been used extensively in general cell culture applications to study cell attachment, proliferation, and invasion. It is ideal for cultures that require limited reduced growth factor conditions.
Key Benefits:	Ideal for general cell culture application Reduced growth factor formulation provides a more defined culture system Polymerizes at 37°C to form a reconstituted basement membrane Quality controlled for peformance consistency

Technical Data
Citations / Reviews (8) / FAQs

Summary for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Why Use Cultrex RGF BME?

Cultrex Reduced Growth Factor Basement Membrane Extract (RGF BME) is a soluble form of basement membrane purified from Engelbreth-Holm-Swarm (EHS) tumor. It is processed to reduce matrix-associated growth factors to provide a matrix that can be used to create a more defined model system. This extract provides a natural extracellular matrix hydrogel that polymerizes at 37°C to form a reconstituted basement membrane. Cultrex RGF BME can be used in multiple applications, including maintaining growth or promoting differentiation of primary endothelial, epithelial, smooth muscle, stem cells, and organoid/3-D cell cultures. It can also be utilized in cell attachment, neurite outgrowth, angiogenesis, in vitro cell invasion, and in vivo tumorigenicity assays.

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.

Product Specifications for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Species	Mouse
Source	Murine Engelbreth-Holm-Swarm (EHS) tumor
Protein Concentration	8-12 mg/mL as determined by Lowry assay
Endotoxin level	≤ 8 EU/mL by Limulus Amoebocyte Lysate (LAL) assay
Sterility Testing	No bacterial or fungal growth detected following 14 days in culture
Cell Culture Testing	Tube Formation Assay - Cultrex RGF BME supports formation of capillary-like structures by human (HBMVEC; HUVEC) or mouse (SVEC4-10) endothelial cells in a tube formation assay.
Viral Testing	Tested negative by PCR test for a total of 31 organisms and viruses, including: mycoplasma, 17 bacterial and virus strains typically included in mouse antibody production (MAP) testing, and 13 additional murine infectious agents including LDEV.

Scientific Data Examples for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

	Assessment of Pluripotency on Cultrex RGF BME. Alkaline Phosphatase (AP) staining was used to characterize the pluripotency of human induced pluripotent stem (iPS) cells cultured on Cultrex Reduced Growth Factor BME (Catalog # 3433-010-01). Images are shown in 4X and 10X magnification. Data courtesy of GABAeron.

Preparation & Storage

Stability	Product is stable for at least two years from date of manufacture when stored at ≤ -70 °C. See lot specific Certificate of Analysis for expiration date.
Shipping Conditions	The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended on the product label.
Storage	Store the unopened product at -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear Best Seller

R&D Systems, part of Bio-Techne | Catalog # 3433-010-01

Key Product Details

Summary for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Why Use Cultrex RGF BME?

Product Specifications for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Scientific Data Examples for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Preparation & Storage

FAQs for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Product Documents for Cultrex Reduced Growth Factor Basement Membrane Extract, PathClear

Product Datasheets

COA

Certificate of Analysis

SDS

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