BD Matrigel™ Basement Membrane Matrix and BD Matrigel™ Basement Membrane Matrix High Concentration (HC) Cells Behave Better on BD Matrigel™ Matrix BD Matrigel™ Matrix is a solubulized basement membrane preparation extracted from EHS mouse sarcoma, a tumor rich in ECM proteins. Its major component is laminin, followed by collagen IV, heparan sulfate proteoglycans, and entactin 1. At room temperature, BD Matrigel™ Matrix polymerizes to produce biologically active matrix material resembling the mammalian cellular basement membrane. Cells behave as they do in vivo when they are cultured on BD Matrigel™ Matrix. It provides a physiologically relevant environment for studies of cell morphology, biochemical function, migration or invasion, and gene expression. BD Matrigel™ Matrix - The trusted name in life science research and drug discovery for over 15 years. Lot to lot consistency Quality assurance testing Reproducibility Wide Selection: BD Matrigel™ Matrix Growth Factor Reduced (GFR) is suited for applications where a more highly defined basement membrane preparation is desired BD Matrigel™ Matrix Phenol-Red Free format is recommended for assays which require color detection (i.e. fluorescence) BD Matrigel™ Matrix HC is suited for in vivo applications where a high protein concentration augments growth of tumors. The high protein concentration also allows the BD Matrigel™ Matrix Plug to maintain its integrity after subcutaneous injection into mice.
Applications
Cell Growth and Differentiation BD Matrigel™ Matrix is especially suited for the culture of polarized cells, such as epithelial cells. It promotes the differentiation of many cells types, including hepatocytes, mammary epithelial, endothelial, smooth muscle cells and neurons 1-8. The acini shown were stained with H/E at 72 hours. Indirect immunofluorescence staining also revealed the salivary gland specific cysteine protease inhibitor, cystatin, in HSG cell acini (data not shown). (Photo courtesy of Dr. Hynda Kleinman).   Metabolism/Toxicology Studies BD Matrigel™ Matrix has been used to successfully construct in vitro models of liver cells for drug toxicity studies 9-13. Note the clusters of spherical cells for hepatocytes cultured on BD Matrigel Matrix, typical of differentiated cells.   Invasion Assays BD Matrigel™ Matrix provides a biologically active basement membrane model for in vitro invasion assay 14-19. Scanning electron micrograph of two human fibrosarcoma cells, having digested the BD Matrigel Matrix occluding the membrane and migrating through the 8 µm of the PET membrane.   In Vitro and In Vivo Angiogenesis Assays BD Matrigel™ Matrix serves as a substrate for in vitro endothelial cell invasion and tube formation assays. It can also be used to assess in vivo angiogenic activity of different compounds via the BD Matrigel Plug Assay 20-27.   　 In Vivo Angiogenesis Studies and Augmentation of Tumors in Immunosuppressed Mice BD Matrigel™ Matrix HC (NEW) is suited for in vivo applications where a high protein concentration augments growth of tumors. The high protein concentration also allows the BD Matrigel Matrix Plug to maintain its integrity after subcutaneous injection into mice. This keeps the injected tumor cells and/or angiogenic compounds localized for in situ analysis and/or future excision 28-41.
Characterization
Amounts of Growth Factors (GF) Present in BD Matrigel™ Matrix vs. GFR BD Matrigel™ Matrix Growth Factor Range of GF Concentration in BD Matrigel Matrix Average GF Concentration in BD Matrigel Matrix Typical GF Concentration in GFR BD Matrigel Matrix EGF 0.5-1.3 ng/ml 0.7 ng/ml < 0.5 ng/ml bFGF < 0.1-0.2 pg/ml n.a.* n.d.** NGF < 0.2 ng/ml n.a.* < 0.2 ng/ml PDGF 5-48 pg/ml 12 pg/ml < 5 pg/ml IGF-1 11-24 ng/ml 16 ng/ml 5 ng/ml TGF-β 1.7-4.7 ng/ml 2.3 ng/ml 1.7 ng/ml * n.a. - not applicable **n.d. - not determined ECM Composition of BD Matrigel™ Matrix vs. GFR Matrigel Matrix Basement Membrane Matrigel Component Percent in BD Matrigel™ Matrix Percent in GFR BD Matrigel™ Matrix Laminin 56 61 Collagen IV 31 30 Entactin 8 7
Quality Control
Tested for ability to gel quickly and maintain this form with culture medium for a period of 14 days at 37° C Tested for ability to promote neurite outgrowth of chick dorsal root ganglia cells Tested and found negative for bacteria, fungi, and mycoplasma Tested for endotoxin (LAL assay) BD Matrigel™ Basement Membrane Matrix HC is tested for its ability to gel quickly and maintain this form with culture media for a period of 12 days at 37° C
Source
Engelbreth-Holm-Swarm mouse tumor
Instructions for Use
BD Matrigel™ Matrix may be used undiluted to culture cells on thin (0.5 mm) or thick (1.0 mm) layers. It may also be diluted with serum-free culture medium and applied as a thin layer to culture substrates. Coating concentration will vary depending on cell type and application. Please contact our Technical Services Department for details on preparing Matrigel HC and/or Matrigel HC/Cell Suspensions for injection into mice.
Storage & Stability:
Stable for at least three months when kept frozen at -20° C or 12 days at 37° C. Avoid multiple freeze-thaws. Do not store in frost-free freezer.
Technical Documents / References
BD Matrigel™ Basement Membrane Matrix--Product Specification Sheet [ PDF ] BD Matrigel™ Matrix High Concentration--Product Sheet [ PDF ] The following Technical Notes are available for download or by calling Technical Services at 800.343.2035 Note No. Author/Title 455 Methods for Implantation of BD Matrigel™ Matrix into Mice and Tissue Fixation Kazuo Ohashi, M.D., Ph.D., Takashi Yokoyama, M.D., Yoshiyuki Nakajima, M.D., Ph.D., and Marshall Kosovsky, Ph.D. Introduction Kleinman, H.K., et. al., Biochem. 21:6188 (1982). Cell Growth and Differentiation Kleinman, H.K., Et al., Biochemistry 25:312 (1986). Hadley, M.A., et al., J.Cell.Biol. 101:1511 (1985). Kubota, et al., J.Cell Biol. 107:1589 (1988). McGuire and Orkin, Biotechniques 5#6:456 (1987). Li, M.L., et al., Proc.Nat.Acad.Sci.USA 84:136 (1987). Barcellof, M.H., Et al., Development 105:223 (1989). Madison, R., et al., Exp.Neurology 88:767 (1985). Metabolism/Toxicology Studies Rana, B., et al., Mol. Cell. Bio. 14:5858 (1994). Schuetz, E.G., et al., J. Cell. Physiol. 134:309 (1988). Schuetz, J. D. and Schuetz, E.G., Cell Growth and Diff. 4:31 (1993). Kane, R.E., et al., In Vitro Cell Dev. Biol. 27A:953 (1991). Mann, D. J., et al., J. Mol. Endocrin. 8:235 (1992). Invasion Assays Liotta, L.A., et al., Cancer Metastasis Rev. 1:277 (1982). Melchiori, A., et al., Cancer Res. 52:2353 (1992). Taniguchi, B.M., et al., Cancer Research 49:6738 (1989). Terranova, V.P., et al., Proc. Nat. Acad. Sci. USA 83:465 (1986). Albini, A., et al., Cancer Research 47:3239, (1987). Hendrix, M., et al., Cancer Letters 38:137 (1987). In Vitro Angiogenesis Assay Nicosia, R.F. and Ottinetti, A., In Vitro Cell. Dev. Biol. 26:119 (1990). Furcht, L.T., Lab Invest. 55:505. (1986). Liotta, LA, et al., Cell. 64:327. (1991). Hirata, K., et al., J. Biol. Chem. 274(20):14170 (1999). Kubota, Y., et al., J. Cell. Bio. 107:1589 (1988). Senger, D. R., et al., PNAS 94:13612 (1997). Passanati, A., et al., Lab Invest. 67:519 (1992). Kibbey, M.C., Et al., J. Natl. Canc. Inst. 84:1633 (1992). In Vivo Angiogenesis Assays and Augmentation of Tumors in Immunosuppressed Mice Senger, D.R., et al., PNAS 94:13612 (1997). Kibbey, M.C., et al., J. Natl. Canc. Inst. 84:1633 (1992). Janowska-Wieczorek, A., et. al., Leukemia 16:1160 (2002). Riccioni, T., et. al., Gene Therapy 5:747 (1998). Bandyopadhyay, A., et. al., Oncogene 21:3541 (2002). Wen, R., et. al., Invest. Ophthalmol Vis SCI 43(12):1297 (2002). Pretlow, T.G., et. al., Cancer Res. 51:3841 (1991). Noel, A., et. al., Biochem. Pharmacol. 43:1263 (1992). Mehta, R.R., et. al., Breast Cancer Res. and Treatment 25:65 (1993). Fridman, R., et. al., J. National Cancer Inst. 83:769 (1991). Sterling-Levis, K., et. al., Cancer Res. 53:1222 1(1993). Noel, A., et. al., Anticancer Res. 15:1 (1995). Fridman, R., et. al., J. National Cancer Inst. 83(11):769 (1991). Glondu, M., et. al., Oncogene 20:6920 (2001).
Ordering Information
Catalog Number Description Qty/Pack BD Matrigel™ Basement Membrane Matrix * 356234 Formulation: Dulbeccos Modified Eagles Medium with 50 µg/ml gentamycin. BD Matrigel™ Matrix is compatible with all culture media. 5 ml 354234 10 ml 356235 50 ml 5x10 ml) BD Matrigel™ Basement Membrane Matrix High Concentration 354248 Formulation: Dulbeccos Modified Eagles Medium with 50 µg/ml gentamycin. BD Matrigel™ Matrix HC is compatible with all culture media. 10 ml BD Matrigel™ Matrix , Phenol Red-free* 356237 Formulation: Dulbeccos Modified Eagles Medium (without phenol red) with 50 µg/ml gentamycin. Phenol Red-free BD Matrigel™ Matrix is compatible with all culture media. 10 ml Growth Factor Reduced (GFR) BD Matrigel™ Matrix*† 356230 Formulation: Dulbeccos Modified Eagles Medium with 50 µg/ml gentamycin. GFR BD Matrigel™ Matrix is compatible with all culture media. 5 ml 354230 10 ml Growth Factor Reduced BD Matrigel™ Matrix, Phenol Red-free*† 356231 Formulation: Dulbeccos Modified Eagles Medium (without phenol red) with 50 µg/ml gentamycin. Phenol Red-free BD Matrigel™ Matrix is compatible with all culture media. 10 ml †Purified by the method of Taub et al., reducing the level of heparan sulfate proteoglycan and several growth factors (e.g., EGF, bFGF, IGF-1, PDGF, and NGF, but not TGF-β).
Tips
The use of BD Cell Recovery Solution or Dispase is necessary to recover cells cultured on BD Matrigel™ Matrix. For biochemical analyses (mRNAs or proteins), use BD Cell Recovery Solution (Cat. No. 354253) for recovering cells from BD Matrigel Matrix. Use Dispase (Cat. No. 354235) when single cell suspensions are required for cell counting-replating. Prior to preparation of gel, thaw BD Matrigel™ Matrix at 4°C on ice overnight. Keep on ice until use. When preparing gel, use precooled pipettes, plates, and tubes. BD Matrigel™ Matrix will gel rapidly at 22-35°C. (Caution: Gel may liquify if stored for several hours at 4°C).