Matrigel basement matrix

Manufactured by BD

Sourced in United States, Germany

Matrigel basement matrix is a gelatinous protein mixture extracted from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is primarily composed of laminin, collagen IV, heparan sulfate proteoglycans, and entactin. This complex mixture mimics the extracellular matrix found in many tissues and is commonly used as a substrate for cell culture and tissue engineering applications.

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Close spelling variants of this product

(GFR) Matrigel Basement Membrane Matrix Matrigel GFR Basement Membrane Matrix LDEV-Free Matrigel basement membrane matrix growth factor reduced Matrigel basement membrane-like matrix BD Matrigel™ Basement Membrane Matrix High Concentration Basement Matrigel-coated membrane matrix Matrigel Basement Membrane Matrix BD Matrigel basement membrane matrix (Matrigel) Basement Matrigel Matrigel matrix

18 protocols using matrigel basement matrix

Transwell Assay for Cell Migration

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Transwell assays were performed in 12-well polycarbonate transwell migration chambers with 8-μm pores (Corning), according to the instructions. Briefly, the transwell plates were coated with Matrigel Basement Matrix (BD Biosciences). The lower compartment of the chamber was loaded with 700 μl of control medium or medium from 293T cells transfected with the following vectors: pcDNA/CARS2, pEGFP-N1/KAL1, or KAL1 G146T. The NLT cells were grown in complete medium until they reached subconfluence, and 4 × 10⁴ cells were seeded in the upper surface of the Boyden chamber in medium containing 1% serum. After 24 h, the cells were stained with crystal violet. The number of adherent cells was counted after the non-migrated cells were scraped off of the upper surface of the porous filter. The experiments were performed at least in triplicate.

Wang F., Huang G.D., Tian H., Zhong Y.B., Shi H.J., Li Z., Zhang X.S., Wang H, & Sun F. (2015). Point mutations in KAL1 and the mitochondrial gene MT-tRNAcys synergize to produce Kallmann syndrome phenotype. Scientific Reports, 5, 13050.

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Matrigel Invasion Assay of MDA-MB-231 Cells

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The invasion capability was assessed by the Boyden chamber with an 8 µm pore of polyethylene terephthalate polyester membrane coated with the Matrigel Basement Matrix (BD Biosciences, San Jose, CA). A total of 2.5 × 10³ of MDA-MB-231 cells were suspended in FBS-free RPMI-1640 medium containing DMSO vehicle, or 10 µg/mL ANR, ANS, or ANL were seeded into each top insert. Cells were allowed to invade to the bottom of the membrane, which was immersed in 10% FBS medium for 24 hours. The following staining and imaging procedures were the same as the Boyden chamber migration assay.

Kuo C.Y., Weng T.S., Kumar K.J., Tseng Y.H., Tung T.W., Wang S.Y, & Wang H.C. (2019). Ethanol Extracts of Dietary Herb, Alpinia nantoensis, Exhibit Anticancer Potential in Human Breast Cancer Cells. Integrative Cancer Therapies, 18, 1534735419866924.

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Preclinical evaluation of CRAF-fusion models

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Homozygous NSG mice were obtained for Jackson Laboratories, bred in our animal facility and housed under aseptic conditions. We used 6–10 weeks old mice with equal sex representation, randomized for treatment and no investigator blinding. The Children’s Hospital of Philadelphia Institutional Animal Care and Use Committee approved all animal protocols.
Mouse flank xenograft studies with CRAF-fusion expressing NIH3T3: NIH3T3 cell lines were injected subcutaneously into NSG mice flanks (n=10 each cell line, per treatment arm) and tumor growth was measured daily. Trametinib (1 mg/kg/dose) and everolimus (10 mg/kg/dose) combinatorial drug study was performed by pre-treating with daily oral gavage for one week prior to injecting cell lines (doses chosen based on scaling from human dosing). Ellipsoid tumor volume was calculated using the formula: volume = 1/2(length × width²).
Intracranial (IC) tumor model: 1×10⁶ PMAs expressing SRGAP3-RAF1, QKI-RAF1 and vector control were resuspended in Matrigel basement matrix (BD Sciences) and 2 microliter was injected into the right striatum of NSG mice (n=5/cell line). Animals were monitored and sacrificed at the onset of neurological symptoms, QKI-RAF1 at day 39 and SRGAP3-RAF1 at day 42 post-injection. Similar treatments doses as used in flank experiment were started at day 13 post-recovery from IC injection. All animals were sacked at day 50.

Jain P., Fierst T.M., Han H.J., Smith T.E., Vakil A., Storm P.B., Resnick A.C, & Waanders A.J. (2017). CRAF gene fusions in pediatric low-grade gliomas define a distinct drug response based on dimerization profiles. Oncogene, 36(45), 6348-6358.

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Transwell-based Cell Migration and Invasion Assay

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The Transwell-based migration and invasion assays were conducted in the modified Boyden chambers (Transwell; Corning Inc. Lowell, MA, USA) as described previously (53 (link)). When the growing cells reached 70% confluency, they were starved for 12 h in serum-free medium. The experimental cells (5 × 10³) were suspended in 0.5 ml medium containing 5% FBS and seeded in the upper chamber of a Transwell, which allows the cells to grow on the microporous polycarbonate membrane that is tissue culture-treated to enhance the cellular attachment to the bottom. The cell-seeded Transwells were placed in each well of 24-well plates containing 1 ml of complete medium (i.e., the lower chamber), and then cultured for 24 h in the incubator at 37 °C with 5% CO₂. Of note, the bottom of upper Transwell was pre-coated by matrigel basement matrix (BD, Biosciences, USA), before the cells were placed in the invasion assay. The remaining cells in the upper chamber were removed, and the cells attached to the lower surface of the Transwell membranes were fixed with 4% paraformaldehyde (AR10669, BOSTER) and stained with 1% crystal violet reagent (Sigma) before being counted.

Wang M., Ren Y., Hu S., Liu K., Qiu L, & Zhang Y. (2021). TCF11 Has a Potent Tumor-Repressing Effect Than Its Prototypic Nrf1α by Definition of Both Similar Yet Different Regulatory Profiles, With a Striking Disparity From Nrf2. Frontiers in Oncology, 11, 707032.

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Antibody Acquisition and Protein Reagents

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Antibodies against E-cadherin, N-cadherin, and Matrigel basement matrix were purchased from BD Biosciences (San Jose, CA, USA). Antibodies against IGFBP-3, vimentin, myc, glutathione S-transferase (GST), His-probe, HA-probe, OctA-probe, IGF-1R, ubiquitin, and actin were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Antibodies against E-cadherin, N-cadherin, and tubulin were purchased from Cell Signaling Technology (Danvers, MA, USA). Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from GeneTex (Irvine, CA, USA). Fluorochrome (Alexa Fluor 488, Alexa Fluor 546, or Alexa Fluor 594)-conjugated secondary antibodies were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Human recombinant IGFBP-3 was purchased from R&D Systems (Minneapolis, MN, USA). Human recombinant IGFBP-3 protein used or evaluation of cellular uptake of extracellular IGFBP-3 was kindly provided by Insmed Inc. (Glen Allen, VA, USA) [22 (link)]. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from MP Biomedicals (Santa Ana, CA, USA). G418 was purchased from Enzo Life Sciences (Farmingdale, NY, USA). Crystal violet, a mouse monoclonal anti-vimentin antibody, and additional chemicals unless otherwise indicated were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Le H.T., Lee H.J., Cho J., Min H.Y., Lee J.S., Lee S.J, & Lee H.Y. (2021). Insulin-Like Growth Factor Binding Protein-3 Exerts Its Anti-Metastatic Effect in Aerodigestive Tract Cancers by Disrupting the Protein Stability of Vimentin. Cancers, 13(5), 1041.

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Investigating Signaling Pathways in Cancer

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Antibodies (Abs) against MMP-1, MMP-2, COX-2, myeloid cell leukemia-1 (Mcl-1), Bax, Fas, Fas-L, AKT, phosphorylated AKT (p-AKT) (Ser473), survivin, Beclin-1 and light chain 3 (LC3) were purchased from Cell Signaling Technology (Danvers, Massachusetts, USA). Abs against E-cadherin and EP2 were from Abcam (Hong Kong, China). PGE2 protein, recombinant human MMP-2 protein (rh-MMP-2) and meloxicam dissolved in dimethyl sulfoxide (DMSO) were purchased from Merck Millipore (Merck Millipore, Darmstadt, Germany). Matrigel basement matrix (10 mg/ml) was purchased from BD Biosciences (San Jose, CA, USA), MK-2206 (an AKT inhibitor) from Jinan Trio Pharmatech Co., Ltd. (Jinan, China), and two autophagy inhibitors 3-methyladenine (3-MA) and chloroquine (CQ) from Sigma-Aldrich (Shanghai, China).

Dong X., Li R., Xiu P., Dong X., Xu Z., Zhai B., Liu F., Jiang H., Sun X., Li J, & Qiao H. (2014). Meloxicam Executes Its Antitumor Effects against Hepatocellular Carcinoma in COX-2- Dependent and -Independent Pathways. PLoS ONE, 9(3), e92864.

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Transwell-Based Cell Migration and Invasion Assay

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The Transwell-based migration and invasion assays were conducted in the modified Boyden chambers (Transwell; Corning Inc. Lowell, MA, USA). Equal numbers of cells were allowed for growth in each well of 12-well plates. After reaching 70–80% confluence, they were starved for 12 h in a serum-free medium. The experimental cells (1 × 10⁴) were suspended in a 0.5 ml medium containing 5% FBS and seeded in the upper chamber of a Transwell. The cell-seeded Transwells were placed in each well of 24-well plates containing 1 ml of complete medium (i.e., the lower chamber), and then cultured for 24 h in the incubator at 37 °C with 5% CO2. Of note, the bottom of the upper Transwell was pre-coated by matrigel basement matrix (BD, Biosciences, USA), before the cells were placed in the invasion assay. The remaining cells in the upper chamber were removed, and the cells attached to the lower surface of the Transwell membranes were fixed with 4% paraformaldehyde (AR10669, BOSTER) and stained with 1% crystal violet reagent (Sigma) before being counted.

Hu S., Feng J., Wang M., Wufuer R., Liu K., Zhang Z, & Zhang Y. (2022). Nrf1 is an indispensable redox-determining factor for mitochondrial homeostasis by integrating multi-hierarchical regulatory networks. Redox Biology, 57, 102470.

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Subcutaneous Xenograft Model of Colorectal Cancer

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All animal studies were performed with the approval of the Austin Health Animal Ethics Committee. 8-week-old male Balb/c nu/nu mice were obtained from the Animal Resources Centre (ARC, Perth, Australia) and maintained in specific pathogen free (SPF) microisolators. SW1116, SW403, SW948, T84, RKO, HCT116, LIM2405, Colo320, HCT116^EV, HCT116^EHF, HCT116^CDX1 and HCT116^{EHF + CDX1}, SW948-NT, SW948-siEHF, SW948-siCDX1 and SW948-siEHF+siCDX1 (2 × 10⁶ cells) CRC cell lines were injected subcutaneously into the left and right flank of each animal in a 150 µL suspension of 1:1 mixture of DMEM:Matrigel Basement Matrix (BD Sciences). Ten mice were used per isogenic cell line. Tumour growth was monitored three times per week by caliper measurement and tumour volume calculated according to the formula pi/6 × large diameter × small diameter². Once the tumours reached a size of 1 cm³, animals were euthanized, and tumours were fixed in 10% formalin and paraffin embedded.

Luk I.Y., Jenkins L.J., Schoffer K.L., Ng I., Tse J.W., Mouradov D., Kaczmarczyk S., Nightingale R., Burrows A.D., Anderson R.L., Arango D., Dopeso H., Croft L., Richardson M.F., Sieber O.M., Liao Y., Mooi J.K., Vukelic N., Reehorst C.M., Afshar-Sterle S., Whitehall V.L., Fennell L., Abud H.E., Tebbutt N.C., Phillips W.A., Williams D.S., Shi W., Mielke L.A., Ernst M., Dhillon A.S., Clemons N.J, & Mariadason J.M. (2022). Epithelial de-differentiation triggered by co-ordinate epigenetic inactivation of the EHF and CDX1 transcription factors drives colorectal cancer progression. Cell Death and Differentiation, 29(11), 2288-2302.

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Transwell Migration and Invasion Assay

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Both Transwell migration and invasion assays were performed in modified Boyden chambers (Transwell; Corning Inc. Lowell, MA, USA) as described previously69 (link). Briefly, after the growing cells reached 70% confluency, they were starved for 12 h in a serum-free medium before being tripsinised. Then experimental cells (5 × 10³) were suspended in 0.5 ml medium containing 5% FBS and seeded in the upper chamber of a Transwell, which allows the cells to grow on the microporous polycarbonate membrane that is tissue culture-treated to enhance cell attachment to the bottom, after migratory cells passed through the 8-μm microporous membrane. The cell-seeded Transwells were placed in each well of 24-well plates containing 1 ml complete medium (i.e. the lower chamber), and then cultured for 24 h in the incubator at 37 °C with 5% CO₂. Of note, the upper chamber bottom of Transwells was pre-coated by matrigel basement matrix (BD, Biosciences, USA) before the cells were placed in the invasion assay. The remaining cells in the upper chamber were removed, and then the cells attached to the lower surface of the Transwell membranes were fixed with 4% paraformaldehyde and stained with 1% crystal violet reagent before being counted.

Ren Y., Qiu L., Lü F., Ru X., Li S., Xiang Y., Yu S, & Zhang Y. (2016). TALENs-directed knockout of the full-length transcription factor Nrf1α that represses malignant behaviour of human hepatocellular carcinoma (HepG2) cells. Scientific Reports, 6, 23775.

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Migration and Invasion Assay Protocol

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Migration assays were performed using 24-well Transwell chambers with polyethylene terephthalate (PET) membrane of 8 μm pore size (BD Biosciences, MA, USA). Cells were serum-starved for 18 hours and detached with Trypsin-EDTA (Gibco, Auckland, NZ). Then, 1×10⁵ cells were suspended in serum-free medium and added to the upper chambers of the Transwell inserts and 3T3 cell-conditioned medium or medium containing 1U/ml thrombin was used as chemoattractant in the lower chamber. After 48 hours of incubation at 37°C, cells in the upper surface of the membranes were removed with a cotton swab. The membranes were fixed with 4% formaldehyde for 15 minutes and stained with 0.2% crystal violet for 10 minutes, and further rinsed with water. The membranes were detached from the inserts and mounted onto glass slides using DPX mountant. Stained cells were counted in 4 randomly chosen microscopic fields per insert at 200x magnification and average value of 3 biological replicates was obtained.
Cell invasion was assayed with Biocoat Matrigel 24-well invasion chamber (BD Biosciences, MA, USA) according to the method described for the Transwell migration assay, with the exception that inserts were pre-coated with matrigel basement matrix (BD Biosciences, MA, USA).

Gan C.P., Patel V., Mikelis C.M., Zain R.B., Molinolo A.A., Abraham M.T., Teo S.H., Abdul Rahman Z.A., Gutkind J.S, & Cheong S.C. (2014). Heterotrimeric G-protein alpha-12 (Gα12) subunit promotes oral cancer metastasis. Oncotarget, 5(20), 9626-9640.

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