BD Matrigel™ Basement Membrane Matrix and BD Matrigel™ Basement Membrane Matrix High Concentration (HC)

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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: Dulbecco's Modified Eagle's 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: Dulbecco's Modified Eagle's 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: Dulbecco's Modified Eagle's 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: Dulbecco's Modified Eagle's 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: Dulbecco's Modified Eagle's 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).

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BD Matrigel™ Basement Membrane Matrix and BD Matrigel™ Basement Membrane Matrix High Concentration (HC)

Cells Behave Better on BD Matrigel™ Matrix