Matrigel

Manufactured by Corning

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Matrigel is a complex mixture of extracellular matrix proteins derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is widely used as a basement membrane matrix to support the growth, differentiation, and morphogenesis of various cell types in cell culture applications.

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Lab products found in correlation

Close spelling variants of this product

Matrigel Basement Membrane Matrix (371 citations) Matrigel Growth Factor Reduced Basement Membrane Matrix (118 citations) PBS/Matrigel (36 citations) Matrigel-coated chambers (32 citations) Matrigel-coated wells (13 citations) Matrigel glue (13 citations) Precooled Matrigel (6 citations) BioCoat Matrigel Invasion Chamber 24-well plate 8.0 μm (5 citations) Matrigel™ GFR Membrane Matrix (5 citations) Growth factor reduced/phenol free matrigel (4 citations)

8 653 protocols using matrigel

Comprehensive T Cell Infiltration Analysis

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RencaHA tdTomato cells were resuspended at a concentration of 1 × 10⁵ cells/ml, mixed with Matrigel (Corning) at 4 °C, seeded in a 24-well plate at a final concentration of 500 cells per Matrigel dome, and left to solidify for 10 min at 37 °C. 2 ml cell medium was added to each well and cells incubated at 37 °C for 11 days. Each Matrigel dome was washed twice in PBS and incubated for 30 min with 1 ml of Cell Recovery Solution (Corning). Spheroids were collected in a 15-ml Falcon tube and pulsed with K^dHA peptide at a final concentration of 2 μg/ml for 1 h. Pulsed spheroids were re-embedded in Matrigel together with 5 × 10⁵ primed CL4 CTL per Matrigel dome. Matrigel domes were dissolved for analysis of spheroid-infiltrating T cells after 16 h: Spheroids were washed twice in PBS and incubated with 1 ml of Cell Recovery Solution (Corning). Spheroids were collected, washed through a 70 μm sieve, and then disaggregated to retrieve T cells in 500 μl of imaging buffer for immediate FACS sorting.

Edmunds G.L., Wong C.C., Ambler R., Milodowski E.J., Alamir H., Cross S.J., Galea G., Wülfing C, & Morgan D.J. (2022). Adenosine 2A receptor and TIM3 suppress cytolytic killing of tumor cells via cytoskeletal polarization. Communications Biology, 5, 9.

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Transwell Cell Invasion Assay with Matrigel

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Cell invasion assay using 24 wells transwell (pore size 8 μm; Corning) was pre‐coated Matrigel (Falcon 354480; Corning). Matrigel (5 mg/ml) was purchased from Corning and stored in ‐20℃, and before transwell assay, the Matrigel was placed in 4℃ overnight and diluted in RPMI1640 with ratio 1:10; the upper compartment was precoated with 50‐μl diluted Matrigel and incubated at 37℃ for 2 h to allow the Matrigel solidification. A total of 1 × 10⁵ cells were suspended in 500 μl RPMI 1640 with 1% FBS and added to the upper chamber, while 750 μl RPMI 1640 contains 10% Place FBS on the bottom for 48 h. Count and take pictures under five optical microscope fields (×100 magnification). Every experiment is repeated three times. Transendothelial invasion test was functioned to discover GFP‐expressing HCT‐116 cells that were invaded by HUVEC monolayer with or without exosome treatment.

Dou R., Liu K., Yang C., Zheng J., Shi D., Lin X., Wei C., Zhang C., Fang Y., Huang S., Song J., Wang S, & Xiong B. (2021). EMT‐cancer cells‐derived exosomal miR‐27b‐3p promotes circulating tumour cells‐mediated metastasis by modulating vascular permeability in colorectal cancer. Clinical and Translational Medicine, 11(12), e595.

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Spheroid Culture of U87 Glioma Cells

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In brief, the U87 cell line was cultured in T25 culture flasks containing 5 mL of pre-warmed cell culture media (DMEM/F12 + 10% FBS + 1% pen/strep) at 37 °C, 5% CO₂, 95% humidity. The U87 cells at 80% confluency were removed from T25 cell culture flask by trypsinization and suspension of cells were centrifuged at 300×g/10 min. The acquired U87 cells at density of 1 × 10⁴ cells was placed on the surface of well-plate in hanging-drop method and incubated at physiological condition for 48 h to provide desirable U87 spheroid. The culture media was added and incubated for 48 h to provide desirable U87 spheroid. Then, prepared U87 spheroid was removed from surface of well via sterilized blade. Separately, 200 μL matrigel (Corning, matrigel, UK) was poured on surface of well and after 30 min incubation, the prepared U87 spheroid was placed on its surface and in following additional 200 μL matrigel poured on prepared matrix surface which resulted U87 spheroid encapsulation in 3 D model of matrigel.

Nooshabadi V.T., Khanmohammadi M., Shafei S., Banafshe H.R., Malekshahi Z.V., Ebrahimi-Barough S, & Ai J. (2020). Impact of atorvastatin loaded exosome as an anti-glioblastoma carrier to induce apoptosis of U87 cancer cells in 3D culture model. Biochemistry and Biophysics Reports, 23, 100792.

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In-Vitro and In-Vivo Angiogenic Characterization

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RFP-expressing HUVECs (Angio-Proteomie) were used for in-vitro angiogenic characterization. Briefly, basement membrane extract (reduced growth factor, Cultrex) was diluted to 10 mg/mL in sterile DPBS. 70 μL of the diluted basement membrane extract was added to each 96 well and allowed to gel at 37 °C for approximately 1 hour. 15,000 cells were then added to each well; EBM was used in the negative control while growth factor-supplemented EBM (EGM) was used for all other groups (n=4). After 12 hours of incubation, cells were examined every hour to determine the optimal end-point. At 15 hours post-seeding, cells were treated with Hoecst (1 μg/mL) for an additional 30 minutes, washed with DPBS, and imaged. To test the effect of B-PED on angiogenesis in-vivo, 1 mL of Matrigel (Corning) containing the appropriate treatment was injected into the caudal ventral area of Sprague-Dawley rats (negative: only Matrigel, positive: Matrigel + 1.5 μg FGF-2, B-PED: Matrigel + 1.5 μg FGF-2 + 10 mg B-PED). Matrigel plugs were extracted 10 days post-implantation and incubated at 37 °C for 30 minutes in a solution of 4% paraformaldehyde (w/v) and 0.5% glutaraldehyde (w/v). Plugs were then treated with 1 mg/mL sodium borohydride at 37 °C for 4 hours. Samples were embedded in OCT and processed for IF staining as described above.

Hwang M.P., Ding X., Gao J., Acharya A.P., Little S.R, & Wang Y. (2018). A Biocompatible Betaine-functionalized Polycation for Coacervation. Soft matter, 14(3), 387-395.

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Matrigel Sandwich Assay for Lumen Formation

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Matrigel sandwich assays were performed by coating 12-well cell culture plates with 150 μL of Matrigel (Corning, Bedford, MA) and incubated at 37 degrees Celsius for 30 minutes to let the Matrigel solidify. One hundred thousand cells in 500 μL of mammary epithelial basal medium (MEBM) (Lonza, Walkersville, MD) were added to Matrigel coated wells. Cells were allowed to attach to Matrigel for three hours at 37^o C. Medium with floating cells were removed and the bound cells were overlaid with 150 μL of 50% Matrigel and 50% MEBM. After the top layer solidified, 2 mL of MEBM plus bovine pituitary gland extract (BPE) (MEGM SingleQuots, Lonza, Walkersville, MD) were added to the wells and culture medium changed every other day. Lumen formation was assessed after 6 days by light microscopy. Cells were isolated from Matrigel using Cell Recovery Solution by Corning (Bedford, MA) according to manufactures protocol.

Stubblefield K., Chean J., Nguyen T., Chen C.J, & Shively J.E. (2017). The adapter SASH1 acts through NOTCH1 and its inhibitor DLK1 in a 3D model of lumenogenesis involving CEACAM1. Experimental cell research, 359(2), 384-393.

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Organoid RNA-Seq Comparative Protocol

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For RNA-Seq comparisons, organoids were maintained in Corning Matrigel lot no. 1062004 (Matrigel 04) and then plated in either Corning Matrigel (growth factor reduced, phenol red free) lot no. 1062004 (Matrigel 04), Corning Matrigel (growth factor reduced, phenol red free) lot no. 0287001 (Matrigel 01), RnD Cultrex reduced growth factor BME lot no. 1564183 (Cultrex 83), RnD Cultrex reduced growth factor BME lot no. 1586187 (Cultrex 87), or Cultrex UltiMatrix reduced growth factor BME lot no. 1637796 (UltiMatrix 96). Each BME condition was plated in triplicate, and organoids were grown for 3 days before harvesting in TRIzol (Invitrogen) and snap frozen. RNA was isolated using the RNeasy Mini Kit (Qiagen). RNA quality control was performed for all samples using a Qubit Fluorometer (Invitrogen) and TapeStation (RIN greater than 8.5) (Agilent) before RNA-Seq analyses.

Lumibao J.C., Okhovat S.R., Peck K.L., Lin X., Lande K., Yomtoubian S., Ng I., Tiriac H., Lowy A.M., Zou J, & Engle D.D. (2024). The effect of extracellular matrix on the precision medicine utility of pancreatic cancer patient–derived organoids. JCI Insight, 9(1), e172419.

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Cancer Cell Migration and Invasion Assay

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Cells were aliquoted into Transwells chambers (Corning, USA), which were inserted in a 24-well plate. After culturing the cells for 24h, the underside of the polycarbonate membranes were fixed. The cells passing through the membrane were observed under a microscope to evaluate the cancer cell migration. In the invasion test, the Matrigel was placed at the bottom of the chamber before cell inoculation. The Matrigel (Corning, 356234, USA) and the 24-well plate were pre-cooled before the Matrigel was laid and then transferred to a 37°C incubator after the Matrigel was laid evenly.

Zhang A., Zou X., Yang S., Yang H., Ma Z, & Li J. (2023). Effect of NETs/COX-2 pathway on immune microenvironment and metastasis in gastric cancer. Frontiers in Immunology, 14, 1177604.

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Feeder-Free Culture of Human Pluripotent Stem Cells

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For gene targeting, H1 human pluripotent stem cells²⁸ (link) were maintained in DMEM-F12 (Life Technologies cat# 11330-057) supplemented with 20% KSR (Life Technologies cat# 10828-028), 4 ng/mL FGF (Life Technologies cat# PHG0261), 0.1 mM 2-mercaptoethanol (Life Technologies cat #21985-023), 1x L-glutamine (Life Technologies cat# 25030-081), 1x MEM-NEAA (Life Technologies cat# 11140-050), 1x penicillin/streptomycin (Life Technologies cat# 15140-122). Cells were passaged with 1 mg/mL collagenase IV (Life Technologies cat# 17104019) every 4-6 days on mitomycin C inactivated MEF feeders cultured in DMEM medium supplemented with 10% FBS, 1x MEM-NEAA, 1x penicillin/streptomycin, and 1x L-glutamine. For differentiation to β-like cells, H1 cells and targeted H1 cells were adapted for feeder-free conditions by culturing in Matrigel (Corning) coated tissue culture plates in mTeSR (Stem Cell Technology). For coating plates, Matrigel (Corning) was 1:100 diluted in cold DMEM-F12 medium, and 2 mL of diluted Matrigel solution was added to one well of a 6-well plate well, and coat for overnight at room temperature before pre-warming in a 37°C incubator for 1 hour before using. Cells were cultured in mTeSR medium in 6-well plates and passaged very 3-4 days with a 1:4-6 passage ratio.

Ma H., Jeppesen J.F, & Jaenisch R. (2020). Human T Cells Expressing a CD19 CAR-T Receptor Provide Insights into Mechanisms of Human CD19-Positive β Cell Destruction. Cell Reports Medicine, 1(6), 100097.

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Lentiviral Modulation of Placental Explants

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First-trimester placental explants were submerged in DMEM/F12 (Gibco) media and divided into smaller pieces under a dissection microscope under sterile conditions. Pieces containing branching villous-like structures were washed in PBS supplemented with 10% FBS and then subjected to lentiviral treatment. The explants were divided into two groups; one group was incubated with scrambled lentiviral particles, while the other group was incubated with lentiviral particles carrying shRNA for WWTR1 gene knockdown. Both groups were incubated with the respective lentiviral particles for 6 h at 37 °C in a humidified chamber in a 5% CO₂/95% air gas mixture. After 6 h, the explant pieces were rinsed and encapsulated in gel for further culture. For the encapsulation, growth factor reduced Matrigel (Corning) was mixed 1:1 with DMEM/F12 on ice to make a Matrigel suspension; 200 μL of the Matrigel suspension was added to each well of a 24-well plate, and explants were placed centrally and covered with another 200 μL of Matrigel suspension. The plate was then incubated at 37 °C in a humidified chamber in 5% CO₂ for the gel suspension to solidify, thereby encapsulating the explant. Finally, 300 μL EVT medium was added to each well and allowed to culture. EVT medium was changed on days 3 and 5 as mentioned earlier.

Ray S., Saha A., Ghosh A., Roy N., Kumar R.P., Meinhardt G., Mukerjee A., Gunewardena S., Kumar R., Knöfler M, & Paul S. (2022). Hippo signaling cofactor, WWTR1, at the crossroads of human trophoblast progenitor self-renewal and differentiation. Proceedings of the National Academy of Sciences of the United States of America, 119(36), e2204069119.

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Transwell Matrigel Invasion Assay

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Matrigel (Corning Glass Works, Corning, N.Y., USA) was dissolved at 4°C overnight and diluted with 1: 3 serum-free DMEM medium. Then 30 μL of diluted Matrigel (15 μL, 7.5 μL, 7.5 μL, 10 min each) was added to the upper chamber of each Transwell chamber (Corning Glass Works, Corning, N.Y., USA) and Matrigel was evenly spread to cover micro-pores at the bottom of the chamber. The cell suspension was incubated in the upper chamber at a density of 3 × 10⁴ cell/well, and DMEM containing 0.5 ml of 10% FBS was added to the lower chamber. After 48 h, the invasive ability was evaluated by the number of cells penetrating through Matrigel. Five fields were randomly chosen to calculate the cell number. The experiment was performed in triplicate.

Wang Y.J., Zhang Z.F., Fan S.H., Zhuang J., Shan Q., Han X.R., Wen X., Li M.Q., Hu B., Sun C.H., Qiao B., Tao Q., Wu D.M., Lu J, & Zheng Y.L. (2017). MicroRNA-433 inhibits oral squamous cell carcinoma cells by targeting FAK. Oncotarget, 8(59), 100227-100241.

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