BD Extracellular Matrix Proteins
ECM Types
Cells behave better on BD Matrigel™ Matrix.
BD Matrigel™ matrix is effective for the attachment and differentiation of both normal and transformed anchorage-dependent epithelioid and other cell types. These include neurons,1,2 Sertoli cells,3 chick lens,4 vascular endothelial cells,5 and hepatocytes.6 BD Matrigel matrix will influence gene expression in adult rat hepatocytes7 as well as three dimensional culture in mouse8-11 and human12,13 mammary epithelial cells. It will support in vivo peripheral nerve regeneration,14-16 can be used for metabolism and toxicology studies,17,18 and is the basis for several types of tumor cell invasion assays.19,20 BD Matrigel matrix provides the substrate necessary for the study of angiogenesis both in vitro 21,22 and in vivo.23-25 BD Matrigel matrix supports in vivo propagation of human tumors in immunosupressed mice.26-28 BD Matrigel matrix is an effective feeder-free surface for maintenance of human embryonic stem cells29-33 and induced pluripotent stem (hiPS) cells34, which has also been used for neuronal differentiation of hiPS cells.35
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Features
- Trusted provider of extracellular matrices since 1985
- Certified LDEV-Free
- Extensive quality control exceeding industry standard
- Rigorously validated for functionality
- Most extensively referenced 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, phenol-red free format is recommended for assays which require color detection (i.e. fluorescence).
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. This keeps the injected tumor cells and/or angiogenic compounds localized for in situ analysis and/or future excision. Available in standard GFR and phenol-red free formats.
Please see the following information sheet on BD Matrigel High Concentration.
BD Matrigel™ Matrix High Concentration
In Vivo Angiogenesis Studies and Augmentation of Tumors in Immunosuppressed Mice
Fluorochrome-Conjugated Antibodies to Pluripotent Stem Cell Markers for Bioimaging and Flow Cytometric Analysis
hES cells (H9) were cultured in mTeSR™1- maintenance media (Stem Cell Technologies) on BD Falcon™ 96-well imaging plates (cat. no. 353219) that were coated with BD Matrigel™ hESC-qualified matrix (cat. no. 354277). Cells were fixed with 4% paraformaldehyde, followed by BD™ perm/wash buffer (Cat. No. 554723). Multicolor cell staining was performed by incubating cells with the following fluorochrome-conjugated antibodies: Sox2 Alexa Fluor® 647 (cat. no. 560302) pseudo-colored yellow, Oct3/4 Alexa Fluor® 555 (cat. no. 560306) pseudo-colored red, and SSEA-4 Alexa Fluor® 488 (cat. no. 560308) pseudo-colored green. Cell nuclei were counterstained using Hoechst 33342 pseudo-colored blue. Small images on the left show individual antibody staining. The larger panel on the right represents the merged image. The cells were imaged on a BD Pathway™ 435 bioimager using a 10X objective.ECM Composition of BD Matrigel Matrix vs. GFR
BD Matrigel Matrix Component | Percent in
BD Matrigel Matrix | Percent in BD Matrigel Matrix GFR |
|---|---|---|
Laminin | 56% | 61% |
Collagen IV | 31% | 30% |
Entactin | 8% | 7% |
Average Growth Factor Concentrations in BD Matrigel Matrix
Parameter | BD Matrigel Matrix | BD Matrigel Matrix GFR |
|---|---|---|
bFGF (pg/ml) | 0 - 0.1 | n.d. |
EGF (ng/ml) | 0.5 - 1.3 | < 0.5 |
IGF-1 (ng/ml) | 15.6 | 5 |
PDGF (pg/ml) | 12 | < 5 |
NGF (ng/ml) | < 0.2 | < 0.2 |
TGF-β (ng/ml) | 2.3 | 1.7 |
| % Protein that gels | 80 | 83 |
n.d.: not determined | ||
Confocal Image of BD™ HUVEC-2 Cell Tube Formation
BD HUVEC-2 cells were assayed using the BD BioCoat Angiogenesis System: Endothelial Cell Tube Formation. Cells were stained using BD™ Calcein AM. Confocal images were captured using the BD Pathway™ bioimager in confocal mode using the 4X objective (NA 0.13) for quantification of tubule formation.
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 hepatocytes cultured on BD Matrigel matrix, which is typical of differentiated hepatocytes.
Invasion Assays
BD Matrigel matrix provides a biologically active basement membrane model for in vitro invasion assays. Scanning electron micrograph of two human fibrosarcoma cells having digested the BD Matrigel matrix occluding the membrane and migrating through the 8 µm pore 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 matrix plug assay.Frequently Asked Questions
Q: How long does it take to thaw BD Matrigel matrix?
A: The product should be thawed overnight on ice in a 2°C to 6°C refrigerator (in the back), or cold room. It may take longer to thaw if the protein concentration is high.
Q: Do I really need to chill my pipet tips and tubes when using BD Matrigel matrix?
A: Yes. Since BD Matrigel™ matrix will start to form a gel above 10ºC, we recommend the use of pre-cooled pipets, tips, and tubes when handling BD Matrigel matrix.
Q: How long can I store a plate after it is coated with BD Matrigel matrix?
A: It is always better to use BD Matrigel matrix-coated plates the same day, but it is application dependent. Coated plates can be stored in the incubator at 37°C for up to a week in serum-free media.
For answers to other questions, please see the following FAQ Guide.
Quality Control
- Mouse colonies are routinely screened for pathogens via mouse antibody production (MAP) testing
- LDEV-free EHS tumor
- 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 matrix
- Tested and found negative for the presence of bacteria, fungi, and mycoplasma
- Protein concentrations are determined by Lowry method
- Endotoxin units are measured by Limulus Amoebocyte Lysate assay
- BD Matrigel matrix 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
References
Cell Growth and Differentiation
1. Biederer, T. and Scheiffele, P., Mixed-culture assays for analyzing neuronal synapse formation, Nature Protocols, 2(3):670 (2007).
2. 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).
3. 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).
4. Ireland, M.E., Quantification and regulation of mRNAs encoding beaded filament proteins in the chick lens, 16(8):838 (1997).
5. 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).
6. 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).
7. 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).
8. 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).
9. Barcellof, M.H., et al., Functional differentiation and aveolar morphogenesis of primary mammary cultures on reconstituted basement membrane, Development, 105:223 (1989).
10. 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).
11. Xu, R., et al., Extracellular matrix-regulated gene expression requires cooperation of SWI/SNF and transcription factors, J. Biol. Chem., 282(20):14992 (2007).
12. 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).
13. 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).
14. Madison, R., et al., Increased rate of peripheral nerve regeneration using bioresorbable nerve guides and laminin containing gel, Exp. Neurology, 88:767 (1985).
15. 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).
16. 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
17. Bi, Y., et al., Use of cryopreserved human hepatocytes in sandwich culture to measure hepatobiliary transport, Drug Metabo. and Dispos., 34(9):1658 (2006).
18. 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).
19. Invasion Assays 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).
20. 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
21. 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).
22. 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
23. 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).
24. 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).
25. Kisucka, J., et al., Platelets and platelet adhesion support angiogenesis while preventing excessive hemorrhage, Proc. Nat. Acad. Sci. USA, 103(4):855 (2006).
26. Albini, A., et al., Matrigel promotes retinoblastoma cell growth in vitro and in vivo, Int. J. Cancer, 52(2):234 (1992).
27. 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).
28. 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).
hESC and iPSC Culture and Differentiation
29. Xu, C., et al., Feeder-free growth of undifferentiated human embryonic stem cells, Nat. Biotechnol., 19:971 (2001)
30. Amit M., et al., Feeder layer- and serum-free culture of human embryonic stem cells, Biol. Reprod. 70:837 (2004)
31. Ludwig T., et al., Feeder-independent culture of human embryonic stem cells, Nat. Methods, 3:637 (2006)
32. Ludwig T., et al., Derivation of human embryonic stem cells in defined conditions, Nat. Biotechnol., 24(2):185 (2006)
33. Harb N., et al., The Rho-Rock-Myosin signaling axis determines cell-cell integrity of self-renewing pluripotent stem cells, PLoS ONE, 3:e3001 (2008)
34. Takahashi K., Induction of pluripotent stem cells from adult human fibroblasts by defined factors, Cell, 131:861 (2007)
35. Chambers S.M., et al, Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling, Nat. Biotechnol., 27:275 (2009)
