BD Matrigel™ Basement Membrane Matrix

BD Matrigel™ Basement Membrane Matrix is effective for the attachment and differentiation of both normal and transformed anchorage dependent epithelioid and other cell types. These include neurons,^5,6 Sertoli cells,⁷ chick lens,⁸ and vascular endothelial cells,⁹ and hepatocytes.¹⁰ BD Matrigel will influence gene expression in adult rat hepatocytes¹¹ as well as three dimensional culture in mouse^12-15 and human^16,17 mammary epithelial cells. It will support in vivo peripheral nerve regeneration,^18-20 can be used for metabolism and toxicology studies,^21,22 and is the basis for several types of tumor cell invasion assays.^23,24 BD Matrigel provides the substrate necessary for the study of angiogenesis both in vitro ^25,26 and in vivo.^27-29 BD Matrigel also supports in vivo propagation of human tumors in immunosupressed mice.^30-32

Features

• Certified LDEV-Free
  
• Extensive quality control exceeding industry standard
• Rigorously validated for functionality
  
• Most extensively references basement membrane matrix
  

Wide Selection of Basement Membranes

• BD Matrigel Matrix Growth Factor Reduced (GFR) is suited for applications where a more highly defined basement membrane preparation is desired. Available in Standard GFR, High Concentration, and Phenol-Red Free formats.
  
• BD Matrigel Matrix High Concentration (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. Available in Standard, Growth Factor Reduced (GFR) and Phenol-Red Free formats.
  
• BD Matrigel Matrix Phenol-Red Free format is recommended for assays which require color detection (i.e. fluorescence).
• BD Matrigel hESC-Qualified Matrix has been qualified as mTeSR™1-compatible by StemCell Technologies, thus eliminating the need for time-consuming screening, in order to provide the reproducibility and consistency essential for your human embryonic stem (hES) cell research. The mTeSR™1 formulation and BD Matrigel Matrix have been shown to be a successful combination for feeder-free maintenance of different WiCell™ hES cell lines for up to 20 passages. (mTeSR™1 Cat. No. 05850.)

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. 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. 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. 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.

In Vivo Angiogenesis Studies and Augmentation of Tumors in Immunosuppressed Mice BD Matrigel Matrix High Concentration 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. Available in standard Growth Factor Reduced (GFR) and Phenol-Red Free formats.

Characterization

Quality Control

• Mouse colonies are routinely screened for pathogens via Mouse Antibody Production (MAP) testing
• Extensive PCR testing is performed on a number of pathogens, including LDEV, to ensure strict control of raw materials used during the manufacturing of BD Matrigel
• Tested and found negative for bacteria, fungi, and mycoplasma
• Protein concentrations are determined by Lowry method
• Endotoxin units are measured by Limulus Amoebocyte Lysate assay
• BD Matrigel gel stability is tested for a period of 14 days at 37°C
• Biological activity is determined for each lot using a neurite outgrowth assay. Chick dorsal root ganglia are plated on a 1.0 mm layer of BD Matrigel Matrix and must generate positive neurite outgrowth response after 48 hours without addition of nerve growth factor

Technical Documents

References

1. Introduction
   
   
2. Kleinman, H.K., et al., Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma, Biochemistry, 21:6188 (1982).
3. Kleinman, H.K., et al., Basement membrane complexes with biological activity, Biochemistry, 25:312 (1986).
4. Vukicevic, S., et al., Identification of multiple active growth factors in basement membrane Matrigel suggests caution in interpretation of cellular activity related to extracellular activity related to extracellular matrix components, Experimental Cell Research, 202:1 (1992).
5. McGuire, P.G. and Seeds, N.W., The interaction of plasminogen activator with a reconstituted basement membrane matrix and extracellular macromolecules produced by cultured epithelial cells, J. Cell. Biochem., 40:215 (1989).
   
   Cell Growth and Differentiation
   
   
6. Biederer, T. and Scheiffele, P.,  Mixed-culture assays for analyzing neuronal synapse formation, Nature Protocols, 2(3):670 (2007).
7. Li, Y., et al., Essential Role of TRPC channels in the guidance of nerve growth cones by brain-derived neurotrophic factor, Nature, 434:894 (2005).
8. Hadley, M.A., et al., Extracellular matrix regulates sertoli cell differentiation, testicular cord formation, and germ cell development in vitro, J. Cell Biol., 101:1511 (1985).
9. Ireland, M.E., Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens, 16(8):838 (1997).
10. McGuire, P.G., and Orkin, R.W., A simple procedure to culture and passage endothelial cells from large vessels of small animals, Biotechniques, 5(6):456 (1987).
11. Bissel, D.M., et al., Support of cultured hepatocytes by a laminin-rich gel. Evidence for a functionally significant subendothelial matrix in normal rat liver, J. Clinical Invest., 79:801 (1987).
12. Page, J.L., et al., Gene expression profiling of extracellular matrix as an effector of human hepatocyte phenotype in primary cell culture, Toxilogical Sciences, 97(2):384 (2007).
13. Li, M.L., et al., Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells, Proc. Nat. Acad. Sci. USA, 84:136 (1987).
14. Barcellof, M.H., et al., Functional differentiation and aveolar morphogenesis of primary mammary cultures on reconstituted basement membrane, Development, 105:223 (1989).
15. Roskelley, C.D., et al., Extracellular matrix-dependent tissue-specific gene expression in mammary epithelial cells requires both physical and biochemical signal transduction, Proc. Nat. Acad. Sci. USA, 91(26):12378 (1994).
16. Xu, R., et al., Extracellular matrix-regulated gene expression requires cooperation of SWI/SNF and transcription factors, J. Biol. Chem., 282(20):14992 (2007).
17. Debnath, J., et al., Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures, Methods, 30(3):256 (2003).
18. Muthuswamy, S.K., et al., ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini, Nat. Cell Biol., 3(9):785 (2001).
19. Madison, R., et al., Increased  rate of peripheral nerve regeneration using bioresorbable nerve guides and laminin containing gel, Exp. Neurology, 88:767 (1985).
20. Xu, X.M., et al., Axonal regeneration into Schwann cell-seeded guidance channels grafted into transected adult rat spinal cord, J. Comp. Neurol., 351(1):145 (1994).
21. Fouad, K., et al., Combining schwann cell bridges and olfactory-ensheathing glia grafts with chondroitinase promotes locomotor recovery after complete transection of the spinal cord, The Journal of Neuroscience, 25(5):1169 (2005).
    
    Metabolism/Toxicology Studies
    
    
22. Bi, Y., et al., Use of cryopreserved human hepatocytes in sandwich culture to measure hepatobiliary transport, Drug Metabo. and Dispos., 34(9):1658 (2006).
23. Yu, X., et al., Essential role of extracellular matrix (ECM) overlay in establishing the functional integrity of primary neonatal rat sertoli cell/gonocyte co-cultures: An improved in vitro model for assessment of male reproductive toxicity, Toxilogical Sciences, 84(2):378 (2005).
    
    Invasion Assays
    
    
24. Terranova, V.P., et al., Use of a reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells, Proc. Nat. Acad. Sci. USA, 83:465 (1986).
25. Albini, A., et al., A rapid in vitro assay for quantitating the invasive potential of tumor cells, Cancer Research, 47:3239 (1987).
    
    In Vitro Angiogenesis Assay
    
    
26. Kubota, Y., et al., Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures, J. Cell Biol., 107:1589 (1988).
27. Maeshima, Y., et al., Identification of the anti-angiogenic site within vascular basement membrane-derived Tumstatin, J. Biol. Chem., 276(18):15240 (2001).
    
    In Vivo Angiogenesis Assays and Augmentation of Tumors in Immunosuppressed Mice
    
    
28. Passaniti, A., et al., A simple, quantitative method for assessing angiogenesis and anti-angiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor, Lab Invest., 67:519 (1992).
29. Isaji, M., et al., Tranilast inhibits the proliferation, chemotaxis and tube formation of human microvascular endothelial cells in vitro and angiogenesis in vivo, British Journal of Pharmacology, 122:1061 (1997).
30. Kisucka, J., et al., Platelets and platelet adhesion support angiogenesis while preventing excessive hemorrhage, Proc. Nat. Acad. Sci. USA, 103(4):855 (2006).
31. Albini, A., et al., Matrigel promotes retinoblastoma cell growth in vitro and in vivo, Int. J. Cancer, 52(2):234 (1992).
32. Yue, W., et al., MCF-7 human breast carcinomas in nude mice as a model for evaluating aromatase inhibitors, J. Steroid Biochem. Molec. Biol., 44(4-6):671 (1993).
33. Angelucci, A., et al., Suppression of EGF-R signaling reduces the incidence of prostate cancer metastasis in nude mice, Endocrine-Related Cancer, 13(1):197 (2006).

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