Cultrex 3-D Culture Matrix Reduced Growth Factor Basement Membrane Extract, Pathclear

Reduced Growth Factor Basement Membrane Extract for 3-D Cell Culture
Catalog # Availability Size / Price Qty
3445-001-01
3445-005-01
3445-010-01
Product Details
Citations (13)
FAQs
Reviews (4)

Cultrex 3-D Culture Matrix Reduced Growth Factor Basement Membrane Extract, Pathclear Summary

Cultrex 3-D Culture Matrix RGF BME is an extracellular matrix hydrogel that directs cells to grow in three dimensions and assemble into organotypic structures in vitro.

Key Benefits

• Qualified to support 3-D cell culture applications
• Reduced growth factor formulation provides a more defined culture system
• Quality controlled for performance consistency

Why Use Cultrex 3-D Culture Matrix RGF BME ?

Cultrex 3-D Culture Matrix Reduced Factor Basement Membrane Extract (RGF 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 3-D Culture Matrix RGF BME is produced and qualified specifically for use in 3-D culture studies. It provides the foundation for cells to grow in three dimensions allowing for the formation of complex in vitro structures. To provide the most standardized basement membrane extract for use in 3-D cultures, a special process is employed to reduce growth factors and manufacture matrix at a standard and consistent concentration. This product is then evaluated in 3-D culture assays to validate efficacy. 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.

Specifications

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
Tested following USP <71> sterility guidelines.
Testing Cell Culture
3-D Culture - Cultrex 3-D Culture Matrix RGF BME promotes differentiation of a human epithelial cell line derived from mammary gland (MCF-10A) or human prostate (PC-3) into acinar structures.

Tube Formation Assay - Cultrex 3-D Culture Matrix RGF BME Supports formation of capillary-like structures by human (HBMVEC; HUVEC) or mouse (SVEC4-10) endothelial cells.

Gelling Assay - Cultrex 3-D Culture Matrix RGF BME 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.
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.
Stability
Product is stable for a minimum of 3 months from date of shipment 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.
Species
Mouse

Limitations

For research use only. Not for diagnositic use.

Product Datasheets

Citations for Cultrex 3-D Culture Matrix Reduced Growth Factor Basement Membrane Extract, Pathclear

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. CDK4/6 inhibitors target SMARCA4-determined cyclin D1 deficiency in hypercalcemic small cell carcinoma of the ovary
    Authors: Y Xue, B Meehan, E Macdonald, S Venneti, XQD Wang, L Witkowski, P Jelinic, T Kong, D Martinez, G Morin, M Firlit, A Abedini, RM Johnson, R Cencic, J Patibandla, H Chen, AI Papadakis, A Auguste, I de Rink, RM Kerkhoven, N Bertos, WH Gotlieb, BA Clarke, A Leary, M Witcher, MC Guiot, J Pelletier, J Dostie, M Park, AR Judkins, R Hass, DA Levine, J Rak, B Vanderhyde, WD Foulkes, S Huang
    Nat Commun, 2019;10(1):558.  2019
  2. Impairment of an Endothelial NAD+-H2S Signaling Network Is a Reversible Cause of Vascular Aging
    Authors: A Das, GX Huang, MS Bonkowski, A Longchamp, C Li, MB Schultz, LJ Kim, B Osborne, S Joshi, Y Lu, JH Treviño-Vi, MJ Kang, TT Hung, B Lee, EO Williams, M Igarashi, JR Mitchell, LE Wu, N Turner, Z Arany, L Guarente, DA Sinclair
    Cell, 2018;173(1):74-89.e20.  2018
  3. PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro
    Authors: D Martinez-M, C Jarvis, T Nelius, W de Riese, OV Volpert, S Filleur
    PLoS ONE, 2017;12(4):e0174968.  2017
  4. Tumor-induced pressure in the bone microenvironment causes osteocytes to promote the growth of prostate cancer bone metastases.
    Authors: Sottnik J, Dai J, Zhang H, Campbell B, Keller E
    Cancer Res, 2015;75(11):2151-8.  2015
  5. Survival Outcome and EMT Suppression Mediated by a Lectin Domain Interaction of Endo180 and CD147.
    Authors: Rodriguez-Teja M, Gronau J, Minamidate A, Darby S, Gaughan L, Robson C, Mauri F, Waxman J, Sturge J
    Mol Cancer Res, 0;13(3):538-47.  0
  6. Human mesenchymal stem cells secrete hyaluronan-coated extracellular vesicles.
    Authors: Arasu U, Karna R, Harkonen K, Oikari S, Koistinen A, Kroger H, Qu C, Lammi M, Rilla K
    Matrix Biol, 0;64(0):54-68.  0
  7. Organotypic culture of untransformed and tumorigenic primary mammary epithelial cells.
    Authors: Jechlinger M
    Cold Spring Harb Protoc, 0;2015(5):457-61.  0
  8. AMPK reverses the mesenchymal phenotype of cancer cells by targeting the Akt-MDM2-Foxo3a signaling axis.
    Authors: Chou C, Lee K, Lai I, Wang D, Mo X, Kulp S, Shapiro C, Chen C
    Cancer Res, 0;74(17):4783-95.  0
  9. Silencing beta3 Integrin by Targeted ECO/siRNA Nanoparticles Inhibits EMT and Metastasis of Triple-Negative Breast Cancer.
    Authors: Parvani J, Gujrati M, Mack M, Schiemann W, Lu Z
    Cancer Res, 0;75(11):2316-2325.  0
  10. Tumor-associated macrophages promote invasion while retaining Fc-dependent anti-tumor function.
    Authors: Grugan K, McCabe F, Kinder M, Greenplate A, Harman B, Ekert J, Van Rooijen N, Anderson G, Nemeth J, Strohl W, Jordan R, Brezski R
    J Immunol, 0;189(11):5457-66.  0
  11. In vitro differentiation of human amniotic epithelial cells into insulin-producing 3D spheroids.
    Authors: Okere B, Alviano F, Costa R, Quaglino D, Ricci F, Dominici M, Paolucci P, Bonsi L, Iughetti L
    Int J Immunopathol Pharmacol, 0;28(3):390-402.  0
  12. Live-Cell Imaging of Protease Activity: Assays to Screen Therapeutic Approaches.
    Authors: Chalasani A, Ji K, Sameni M, Mazumder S, Xu Y, Moin K, Sloane B
    Methods Mol Biol, 0;1574(0):215-225.  0
  13. SnoN regulates mammary gland alveologenesis and onset of lactation by promoting prolactin/Stat5 signaling.
    Authors: Jahchan N, Wang D, Bissell M, Luo K
    Development, 0;139(17):3147-56.  0

FAQs

  1. 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.

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

    • Cultrex BME should be stored at or below -20°C for optimal stability. Preparation of working aliquots is recommended. Cultrex BME should be thawed overnight on ice at 4°C, however long term storage at 4°C is not recommended. Freeze/thaw cycles and gel-liquid phase transitions should be avoided, since they can compromise product integrity.

  3. What is the difference between Cultrex® Basement Membrane Extract (BME) and Cultrex® 3D Culture Matrix?

    • Cultrex® 3D Culture Matrix was developed to provide the most standardized basement membrane extract for use in 3D Cultures. A special process is employed to reduce growth factors. This material is then incorporated in a 3D culture to validate efficacy. 3D Culture Basement Membrane Extract promotes differentiation of a human epithelial cell line derived from mammary gland (MCF-10A) and human prostate (PC-3) into acinar structures. The Cultrex® 3D Culture Matrix is essentially the same as our standard Cultrex® BME, but has been additionally qualified with the functional 3D assay as described above.

  4. 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).

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. What type of analysis is typically applied for organoid or 3-D cell cultures?

    • Within the organoid, spheroid, or 3-D culture, cells may be assessed for morphology, apical/basal polarity, protein localization, and relative proliferation. In addition, cells may be isolated from the 3-D culture and evaluated for levels of RNA and protein expression, as well as modifications to DNA.

  11. What are 3-D cultures?

    • 3-D cultures are in vitro cultures where immortalized cell lines, primary cell lines, stem cells, or tissue explants are placed within hydrogel matrices, such as Cultrex Basement Membrane Extract, that mimic in vivo cell environments and allow cells to proliferate in three dimensions.

  12. What is the advantage of 3-D culture over traditional 2-D culture?

    • While 2-D culture has been used for studying many aspects of cell function and behavior, the tissue-culture treated plastic environment is unlike anything found within living organisms. As a result, cells in 2-D culture exhibit altered morphology, function, proliferation, and gene expression when compared to their emanating tissues. By placing these cells in a 3-D environment, they assume biological and biochemical characteristics similar to what is observed in vivo.

  13. What are the variables associated with 3-D culture?

    • The major variables associated with 3-D culture are cell type, cell seeding density, composition of hydrogel, thickness of hydrogel, stiffness of hydrogel, composition of cell culture medium, and time of culture.

  14. What are the different types of 3-D culture?

    • The two principal methods for performing 3-D culture are the top assay and embedded assay. For the top assay, cells are seeded on a thick gel of Cultrex Basement Membrane Extract (BME) or Extracellular Matrix Protein. A thin overlay of cell culture medium is then applied to the cells. For the embedded assay, cells are resuspended within a thick gel of Cultrex BME or ECM and the culture media is applied on top. The top assay is easier to setup, to control seeding densities, and to keep cells within one focal plane for analysis.

  15. Which matrix should I use for 3-D culture?

    • Choice of matrix should correspond to the environment that you wish to recapitulate. Cultrex Basement Membrane Extract (BME) will recapitulate the basal lamina, which underlie most cells of epithelial or endothelial origin. Cultrex Collagen I is the major constituent of connective tissue, and is commonly inhabited by stationary cells, such as fibrocytes and adipose cells, as well as migrating cells, such as mast cells, macrophages, monocytes, lymphocytes, plasma cells, and eosinophils.

  16. How should cells be cultured prior to setting up the 3-D culture?

    • Cells need to be healthy and actively dividing in 2-D culture. Cells should be passaged two or three times after resuspension from cryopreservation, and they should never surpass 80% confluency during each passage. Cells should also be assessed for viability using trypan blue, and they should exhibit less than 5% staining.

  17. What type of plates are recommended?

    • In order for the Cultrex 3-D Culture Matrix RGF BME to stick to the plate, it must be tissue-culture treated.  However, if the application is spheroids or when using BME diluted in media as a slurry to form floating 3-D structures, then low attachment plates are recommended.

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Reviews for Cultrex 3-D Culture Matrix Reduced Growth Factor Basement Membrane Extract, Pathclear

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Cultrex 3-D Culture Matrix Reduced Growth Factor Basement Membrane Extract, Pathclear
By Anonymous on 04/28/2021
Application:

Application: Tumor 3D Proliferation assay


Cultrex 3-D Culture Matrix RGF Basement Membrane Extract
By Anonymous on 01/13/2020
Application: CellProlif

Cultrex 3-D Culture Matrix RGF Basement Membrane Extract
By Anonymous on 10/02/2019
Application: Tumor Spheroids 3D Proliferation assay
Reason for Rating: We used the 3-D culture Cultrex RGF for our Tumor Spheroids 3D Proliferation assay.

Cultrex PathClear 3-D Culture Matrix RGF BME
By Anonymous on 04/01/2018
Application: Apoptosis assay