Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear

For Robust Organoid Cultures - Reduced Growth Factor Basement Membrane Extract
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Purchase Cultrex™ BME with Select Proteins and Save 15% Using Promo Code: PROTBME15. Purchase the 5 or 10 mL size of a select BME product from our catalog along with a minimum of one select growth factor or matrix-regulating protein and receive 15% off your order using promo code: PROTBME15. See offer details for additional qualifying products and terms and conditions.

Mouse Intestinal Organoids Cultured in Cultrex RGF BME Type 2_3533-005-02
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Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear Summary

Cultrex Reduced Growth Factor Basement Membrane Extract (RGF BME ),Type 2 is specifically qualified to support the establishment and expansion of robust organoid cultures. It's composition mimics the in vivo microenvironment to improve take rate and growth of organoids.

Key Benefits

• Qualified for use in organoid cell culture
• Commonly used robust and established organoid systems
• Reduced growth factor formulation provides a more defined culture system
• Quality controlled for performance consistency

Why Use Cultrex RGF Basement Membrane Extract, Type 2?

Cultrex Reduced Growth Factor Basement Membrane Extract (RGF BME), Type 2 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. 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 heparan sulfate proteoglycans.

Cultrex RGF BME, Type 2 provides a proprietary formulation that is high in tensile strength and is designed for use in robust tissue organoid culture as well as other applications requiring an extracellular matrix scaffold.

Protocols Utilizing Cultrex RGF Basement Membrane Extract, Type 2 for Organoid Cell Culture.

Cultrex RGF BME, Type 2 is ideal for use as a scaffold for organoid and 3D cell culture. Listed below are protocols designed by our research and development groups for different types of organoids featuring Cultrex RGF BME, Type 2 as well as growth factors, media supplements, and small molecules from Bio-Techne.
Video of Cultrex BME Best Practices and Protocols.
Protocol for Mouse Enteric Organoid Culture.
Protocol for Human Gastric Organoid Culture.
Protocol for Human Liver Organoid Culture.
Protocol for Human Lung Organoid Culture.
Protocol for Human Intestinal Organoid Culture.
Protocol for Harvesting Organoids for Biochemical Analysis.


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
Testing Cell Culture
Organoid Culture - Cultrex RGF BME, Type 2 supports growth and expansion of mouse small intestine organoid progenitor cells.

Gelling Assay - Cultrex RGF BME, Type 2 gels in less than 30 minutes at 37 °C, and maintains the gelled form in culture medium for a minimum of 7 days at 37 °C.

Dome Assay Cultrex RGF BME, Type 2 forms and maintains stable 3-D dome structures on cell culture plates.

Tube Formation Assay - Cultrex RGF BME, Type 2 supports formation of capillary-like structures by human (HBMVEC; HUVEC) or mouse (SVEC4-10) endothelial cells.
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.
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.
Store the unopened product at -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles.


For research use only. Not for diagnostic use.

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Scientific Data

Mouse Intestinal Organoids Cultured in Cultrex RGF BME, Type 2. Mouse intestinal organoids cultured in Cultrex RGF BME, Type 2 were immersion fixed and processed for whole mount staining or paraffin embedding and sectioning for immunocytochemistry. A) Whole mount mouse intestinal organoids were stained using Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (green; Catalog # AF748) at 10 µg/mL for 3 hours at room temperature. Cells were counterstained with DAPI (blue). B) Paraffin-embedded mouse intestinal tissue stained for Human Cadherin‑17 Antibody (green; Catalog # MAB1032), Human Ki67/MKI67 Antibody (red; Catalog # AF7617), and counterstained with DAPI (blue).

Human Lung Organoids Cultured in Cultrex RGF BME, Type 2. A) Representative brightfield image of human lung organoids cultured using Cultrex RGF BME, Type 2. B) Expression of Sox2 (green; Catalog # AF2018) and Acetylated Tubulin (red; Novus Biologicals, Catalog # NB600-567). C) Expression of p63/TP73L (green; Catalog # AF1916) and Cytokeratin 10 (red; Novus Biologicals, Catalog # NBP2-61736). D) Expression of Podoplanin (green; Catalog # AF3670) as a marker of type 1 alveolar cells.

Liver Organoid Formation and Differentiation in Cultrex RGF BME, Type 2. Human Liver organoids were derived from human biopsy tissue. Undifferentiated organoids were formed by embedding dissociated tissue in Cultrex RGF BME, Type 2 and culturing in specialized media. The organoids were differentiated using media containing Recombinant Human FGF-19 (Catalog # 969-FG), DAPT (Catalog # 2634), and Dexamethasone (Catalog # 1126). A) Undifferentiated liver organoids. B) Liver organoids shrink as they differentiate. C) Representative images of differentiated liver organoids expression hepatocyte markers, Albumin and HNF3 beta, as well as E-Cadherin.

Human Intestinal Organoids Cultured using Cultrex RGF BME, Type 2. Human transverse colon organoids (A,B) and human ileum organoids (C, D) were grown using cells isolated from transverse colon and ileum biopsy tissue, respectively. Organoids were embedded in Cultrex RGF BME, Type 2 as a scaffold matrix. A) Brightfield image of human transverse organoids. B) Human transverse organoid stained using Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (green; Catalog # AF748), a MUC2 Antibody (red; Catalog # NBP2-44431), and DAPI (blue). C) Brightfield image of human ileum organoids. D) Human ileum organoid stained using a Aldolase B Antibody (red; Catalog # NBP2-15345), a Human Cadherin-17 Antibody (green; Catalog # MAB1032, and DAPI (blue).

Formation of HepG2 Spheroids in Cultrex RGF BME, Type 2. Hepatocyte spheroids were formed by plating HepG2 liver hepatocellular carcinoma cells (10,000 cells per well) in a 24-well plate coated with 5 mg/mL of Cultrex RGF BME, Type 2. Spheroids were cultured for 21 days prior imaging by brightfield microscopy.

Citations for Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, 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.

36 Citations: Showing 1 - 10
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  1. Computational pharmacogenomic screen identifies drugs that potentiate the anti-breast cancer activity of statins
    Authors: JE van Leeuwe, W Ba-Alawi, E Branchard, J Cruickshan, W Schormann, J Longo, J Silvester, PL Gross, DW Andrews, DW Cescon, B Haibe-Kain, LZ Penn, DMA Gendoo
    Nature Communications, 2022;13(1):6323.  2022
  2. Division of labor within the DNA damage tolerance system reveals non-epistatic and clinically actionable targets for precision cancer medicine
    Authors: A Spanjaard, R Shah, D de Groot, OA Buoninfant, B Morris, C Lieftink, C Pritchard, LM Zürcher, S Ormel, JJI Catsman, R de Korte-G, B Siteur, N Proost, T Boadum, M van de Ven, JY Song, M Kreft, PCM van den Be, RL Beijersber, H Jacobs
    Nucleic Acids Research, 2022;50(13):7420-7435.  2022
  3. Human branching cholangiocyte organoids recapitulate functional bile duct formation
    Authors: FJM Roos, GS van Tiende, H Wu, I Bordeu, D Vinke, LM Albarinos, K Monfils, S Niesten, R Smits, J Willemse, O Rosmark, G Westergren, DJ Kunz, M de Wit, PJ French, L Vallier, JNM IJzermans, R Bartfai, H Marks, BD Simons, ME van Royen, MMA Verstegen, LJW van der La
    Cell Stem Cell, 2022;29(5):776-794.e13.  2022
  4. RNA splicing is a key mediator of tumour cell plasticity and a therapeutic vulnerability in colorectal cancer
    Authors: AE Hall, SÖ Pohl, P Cammareri, S Aitken, NT Younger, M Raponi, CV Billard, AB Carrancio, A Bastem, P Freile, F Haward, IR Adams, JF Caceres, P Preyzner, A von Kriegs, MG Dunlop, FV Din, KB Myant
    Nature Communications, 2022;13(1):2791.  2022
  5. Cell-intrinsic Aryl Hydrocarbon Receptor signalling is required for the resolution of injury-induced colonic stem cells
    Authors: K Shah, MR Maradana, M Joaquina D, A Metidji, F Graelmann, M Llorian, P Chakravart, Y Li, M Tolaini, M Shapiro, G Kelly, C Cheshire, D Bhurta, SB Bharate, B Stockinger
    Nature Communications, 2022;13(1):1827.  2022
  6. Efficient and error-free fluorescent gene tagging in human organoids without double-strand DNA cleavage
    Authors: Y Bollen, JH Hageman, P van Leenen, LLM Derks, B Ponsioen, JR Buissant d, I Verlaan-Kl, M van den Bo, LWMM Terstappen, R van Boxtel, HJG Snippert
    PloS Biology, 2022;20(1):e3001527.  2022
  7. Comprehensive metabolomics expands precision medicine for triple-negative breast cancer
    Authors: Y Xiao, D Ma, YS Yang, F Yang, JH Ding, Y Gong, L Jiang, LP Ge, SY Wu, Q Yu, Q Zhang, F Bertucci, Q Sun, X Hu, DQ Li, ZM Shao, YZ Jiang
    Cell Research, 2022;0(0):.  2022
  8. Copy number amplification of ENSA promotes the progression of triple-negative breast cancer via cholesterol biosynthesis
    Authors: YY Chen, JY Ge, SY Zhu, ZM Shao, KD Yu
    Nature Communications, 2022;13(1):791.  2022
  9. Lineage-specific silencing of PSAT1 induces serine auxotrophy and sensitivity to dietary serine starvation in luminal breast tumors
    Authors: BH Choi, V Rawat, J Högström, PA Burns, KO Conger, ME Ozgurses, JM Patel, TS Mehta, A Warren, LM Selfors, T Muranen, JL Coloff
    Cell Reports, 2022;38(3):110278.  2022
  10. Air-Liquid-Interface Differentiated Human Nose Epithelium: A Robust Primary Tissue Culture Model of SARS-CoV-2 Infection
    Authors: BM Tran, SL Grimley, JL McAuley, A Hachani, L Earnest, SL Wong, L Caly, J Druce, DFJ Purcell, DC Jackson, M Catton, CJ Nowell, L Leonie, G Deliyannis, SA Waters, J Torresi, E Vincan
    International Journal of Molecular Sciences, 2022;23(2):.  2022
  11. Ultrastructural analysis of breast cancer patient-derived organoids
    Authors: L Signati, R Allevi, F Piccotti, S Albasini, L Villani, M Sevieri, A Bonizzi, F Corsi, S Mazzucchel
    Cancer Cell International, 2021;21(1):423.  2021
  12. ERBB3 overexpression due to miR-205 inactivation confers sensitivity to FGF, metabolic activation, and liability to ERBB3 targeting in glioblastoma
    Authors: F De Bacco, F Orzan, J Erriquez, E Casanova, L Barault, R Albano, A D'Ambrosio, V Bigatto, G Reato, M Patanè, B Pollo, G Kuesters, C Dell'Aglio, L Casorzo, S Pellegatta, G Finocchiar, PM Comoglio, C Boccaccio
    Cell Reports, 2021;36(4):109455.  2021
  13. Adult mouse and human organoids derived from thyroid follicular cells and modeling of Graves' hyperthyroidism
    Authors: J van der Va, L Bosmans, SF Sijbesma, K Knoops, WJ van de Wet, HG Otten, H Begthel, IHM Borel Rink, J Korving, EGWM Lentjes, C Lopez-Igle, PJ Peters, HM van Santen, MR Vriens, H Clevers
    Proceedings of the National Academy of Sciences of the United States of America, 2021;118(51):.  2021
  14. RAC1B modulates intestinal tumourigenesis via modulation of WNT and EGFR signalling pathways
    Authors: V Gudiño, SÖ Pohl, CV Billard, P Cammareri, A Bolado, S Aitken, D Stevenson, AE Hall, M Agostino, J Cassidy, C Nixon, A von Kriegs, P Freile, L Popplewell, G Dickson, L Murphy, A Wheeler, M Dunlop, F Din, D Strathdee, OJ Sansom, KB Myant
    Nature Communications, 2021;12(1):2335.  2021
  15. A Bioluminescent 3CLPro Activity Assay to Monitor SARS-CoV-2 Replication and Identify Inhibitors
    Authors: C Mathieu, F Touret, C Jacquemin, YL Janin, A Nougairède, M Brailly, M Mazelier, D Décimo, V Vasseur, A Hans, JC Valle-Casu, X de Lamball, B Horvat, P André, M Si-Tahar, V Lotteau, PO Vidalain
    Viruses, 2021;13(9):.  2021
  16. Organoid-based drug screening reveals neddylation as therapeutic target for malignant rhabdoid tumors
    Authors: C Calandrini, SR van Hooff, I Paassen, D Ayyildiz, S Derakhshan, MEM Dolman, KPS Langenberg, M van de Ven, C de Heus, N Liv, M Kool, RR de Krijger, GAM Tytgat, MM van den He, JJ Molenaar, J Drost
    Cell Reports, 2021;36(8):109568.  2021
  17. Sox9EGFP defines biliary epithelial heterogeneity downstream of Yap activity
    Authors: DY Tulasi, DM Castaneda, K Wager, CB Hogan, KP Alcedo, JR Raab, AD Gracz
    Cellular and Molecular Gastroenterology and Hepatology, 2021;0(0):.  2021
  18. Culture and analysis of kidney tubuloids and perfused tubuloid cells-on-a-chip
    Authors: L Gijzen, FA Yousef Yen, F Schutgens, MK Vormann, CME Ammerlaan, A Nicolas, D Kurek, P Vulto, MB Rookmaaker, HL Lanz, MC Verhaar, H Clevers
    Nature Protocols, 2021;0(0):.  2021
  19. Inhibition of mitochondrial function by metformin increases glucose uptake, glycolysis and GDF-15 release from intestinal cells
    Authors: M Yang, T Darwish, P Larraufie, D Rimmington, I Cimino, DA Goldspink, B Jenkins, A Koulman, CA Brighton, M Ma, BYH Lam, AP Coll, S O'Rahilly, F Reimann, FM Gribble
    Scientific Reports, 2021;11(1):2529.  2021
  20. Single-cell derived tumor organoids display diversity in HLA class I peptide presentation
    Authors: LC Demmers, K Kretzschma, A Van Hoeck, YE Bar-Epraïm, HWP van den To, M Koomen, G van Son, J van Gorp, A Pronk, N Smakman, E Cuppen, H Clevers, AJR Heck, W Wu
    Nat Commun, 2020;11(1):5338.  2020
  21. An organoid biobank for childhood kidney cancers that captures disease and tissue heterogeneity
    Authors: C Calandrini, F Schutgens, R Oka, T Margaritis, T Candelli, L Mathijsen, C Ammerlaan, RL van Inevel, S Derakhshan, S de Haan, E Dolman, P Lijnzaad, L Custers, H Begthel, HHD Kerstens, LL Visser, M Rookmaaker, M Verhaar, GAM Tytgat, P Kemmeren, RR de Krijger, R Al-Saadi, K Pritchard-, M Kool, AC Rios, MM van den He, JJ Molenaar, R van Boxtel, FCP Holstege, H Clevers, J Drost
    Nat Commun, 2020;11(1):1310.  2020
  22. Slug-expressing mouse prostate epithelial cells have increased stem cell potential
    Authors: Z Kahounová, J Remšík, R Fedr, J Bouchal, A Mi?ková, E Slabáková, L Binó, A Hampl, K Sou?ek
    Stem Cell Res, 2020;46(0):101844.  2020
  23. Functional Radiogenetic Profiling Implicates ERCC6L2 in Non-homologous End Joining
    Authors: P Francica, M Mutlu, VA Blomen, C Oliveira, Z Nowicka, A Trenner, NM Gerhards, P Bouwman, E Stickel, ML Hekkelman, L Lingg, I Klebic, M van de Ven, R de Korte-G, D Howald, J Jonkers, AA Sartori, W Fendler, JR Chapman, T Brummelkam, S Rottenberg
    Cell Rep, 2020;32(8):108068.  2020
  24. Patient-derived oral mucosa organoids as an in vitro model for methotrexate induced toxicity in pediatric acute lymphoblastic leukemia
    Authors: E Driehuis, N Oosterom, SG Heil, IB Muller, M Lin, S Kolders, G Jansen, R de Jonge, R Pieters, H Clevers, MM van den He
    PLoS ONE, 2020;15(5):e0231588.  2020
  25. Organoid cultures from normal and cancer-prone human breast tissues preserve complex epithelial lineages
    Authors: JM Rosenbluth, RCJ Schackmann, GK Gray, LM Selfors, CM Li, M Boedicker, HJ Kuiken, A Richardson, J Brock, J Garber, D Dillon, N Sachs, H Clevers, JS Brugge
    Nat Commun, 2020;11(1):1711.  2020
  26. Preclinical evaluation of the anti-tumor activity of pralatrexate in high-risk neuroblastoma cells
    Authors: RA Clark, S Lee, J Qiao, DH Chung
    Oncotarget, 2020;11(32):3069-3077.  2020
  27. Hypoxia Triggers the Intravasation of Clustered Circulating Tumor Cells
    Authors: C Donato, L Kunz, F Castro-Gin, A Paasinen-S, K Strittmatt, BM Szczerba, R Scherrer, N Di Maggio, W Heusermann, O Biehlmaier, C Beisel, M Vetter, C Rochlitz, WP Weber, A Banfi, T Schroeder, N Aceto
    Cell Rep, 2020;32(10):108105.  2020
  28. Loss of sphingosine 1-phosphate receptor 3 gene function impairs injury-induced stromal angiogenesis in mouse cornea
    Authors: S Yasuda, T Sumioka, H Iwanishi, Y Okada, M Miyajima, K Ichikawa, PS Reinach, S Saika
    Lab Invest, 2020;0(0):.  2020
  29. Pancreatic cancer organoids recapitulate disease and allow personalized drug screening
    Authors: E Driehuis, A van Hoeck, K Moore, S Kolders, HE Francies, MC Gulersonme, ECA Stigter, B Burgering, V Geurts, A Gracanin, G Bounova, FH Morsink, R Vries, S Boj, J van Es, GJA Offerhaus, O Kranenburg, MJ Garnett, L Wessels, E Cuppen, LAA Brosens, H Clevers
    Proc. Natl. Acad. Sci. U.S.A., 2019;0(0):.  2019
  30. Oral mucosal organoids as a potential platform for personalized cancer therapy
    Authors: E Driehuis, S Kolders, S Spelier, K Lohmussaar, SM Willems, LA Devriese, R de Bree, EJ de Ruiter, J Korving, H Begthel, JH Van Es, V Geurts, GW He, RH van Jaarsv, R Oka, MJ Muraro, J Vivie, MMJM Zandvliet, APA Hendrickx, N Iakobachvi, P Sridevi, O Kranenburg, R van Boxtel, GJPL Kops, DA Tuveson, PJ Peters, A van Oudena, H Clevers
    Cancer Discov, 2019;0(0):.  2019
  31. Release of transcriptional repression via ErbB2-induced, SUMO-directed phosphorylation of myeloid zinc finger-1 serine 27 activates lysosome redistribution and invasion
    Authors: DM Brix, SA Tvingsholm, MB Hansen, KB Clemmensen, T Ohman, V Siino, M Lambrughi, K Hansen, P Puustinen, I Gromova, P James, E Papaleo, M Varjosalo, J Moreira, M Jäättelä, T Kallunki
    Oncogene, 2019;0(0):.  2019
  32. SH3BP4 Regulates Intestinal Stem Cells and Tumorigenesis by Modulating ?-Catenin Nuclear Localization
    Authors: P Antas, L Novellasde, A Kucharska, I Massie, J Carvalho, D Oukrif, E Nye, M Novelli, VSW Li
    Cell Rep, 2019;26(9):2266-2273.e4.  2019
  33. Neutrophils escort circulating tumour cells to enable cell cycle progression
    Authors: BM Szczerba, F Castro-Gin, M Vetter, I Krol, S Gkountela, J Landin, MC Scheidmann, C Donato, R Scherrer, J Singer, C Beisel, C Kurzeder, V Heinzelman, C Rochlitz, WP Weber, N Beerenwink, N Aceto
    Nature, 2019;0(0):.  2019
  34. Novel Chimeric Gene Therapy Vectors Based on Adeno-Associated Virus and Four Different Mammalian Bocaviruses
    Authors: J Fakhiri, MA Schneider, J Puschhof, M Stanifer, V Schildgen, S Holderbach, Y Voss, J El Andari, O Schildgen, S Boulant, M Meister, H Clevers, Z Yan, J Qiu, D Grimm
    Mol Ther Methods Clin Dev, 2019;12(0):202-222.  2019
  35. AGE-RAGE interaction in the TGF?2-mediated epithelial to mesenchymal transition of human lens epithelial cells.
    Authors: Cibin T Raghavan, Ram H Nagaraj
    Glycoconjugate Journal, 2016;0(0):1573-4986.  2016
  36. A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity.
    Authors: Sachs N, de Ligt J, Kopper O, Gogola E, Bounova G, Weeber F, Balgobind A, Wind K, Gracanin A, Begthel H, Korving J, van Boxtel R, Duarte A, Lelieveld D, van Hoeck A, Ernst R, Blokzijl F, Nijman I, Hoogstraat M, van de Ven M, Egan D, Zinzalla V, Moll J, Boj S, Voest E, Wessels L, van Diest P, Rottenberg S, Vries R, Cuppen E, Clevers H
    Cell, 0;172(1):373-386.e10.  0


  1. 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.

  2. 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.

  3. 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.

  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. Does Cultrex BME, Catalog # 3533-005-02, affect fluorescence readings when Alamar Blue is used for final readout of assay? 

    • BME is known to have autofluorescence, but if appropriate controls are evaluated, background can be successfully subtracted. A BME only control well with no cells should be used to subtract the background fluorescence.

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Reviews for Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear

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Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear
By Anonymous on 06/18/2022

dissolve it on ice, and don't put it outside for too not time

Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear
By Anonymous on 02/21/2022

Aortic sprouting assay: mouse aortic ring cultured in a 48-well plate and imaged in an inverted microscope after 5 days.

Results are very consistent with this BME (used one layer under and another on top of the aortic ring).

Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear
By Anonymous on 04/13/2021

Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear
By Anonymous on 12/09/2020

We are using the BME at a final dilution of 1/30 for the maintenance of iPSC and differentiation into endoderm as well as mesoderm lineages. Attachment, growth, and differentiation of our iPSC is consistent, and we observed only minor variations between different lots.

Cultrex PathClear Reduced Growth Factor BME (2 x 5 mL)
By wenyi wu on 06/01/2018
Application: Cell migration/motility
Reason for Rating: it can form a very nice tube in our control group within 6-8 hour.