Ecm gel

Manufactured by Merck Group

Sourced in United States, Germany

ECM gel is a complex mixture of extracellular matrix proteins, including collagen, laminin, and proteoglycans. It provides a natural three-dimensional environment for cell culture and tissue engineering applications.

Automatically generated - may contain errors

Lab products found in correlation

Transwell plate, by Corning (6 mentions) Fibronectin, by Merck Group (6 mentions) Transwell chamber, by Corning (5 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Transwell insert, by Corning (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions) Transwell chambers with 8 μm pore, by Corning (2 mentions) Transwell chambers for 24 well plates, by Corning (2 mentions) Collagen 1, by Merck Group (2 mentions) Ckx41, by Olympus (2 mentions) Crystal violet, by Merck Group (2 mentions) Fetal calf serum (fcs), by Merck Group (1 mentions) Alpha mem media, by Merck Group (1 mentions) 96 well plate, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Na2co3, by Merck Group (1 mentions) Human umbilical vein endothelial cells (huvec), by Lonza (1 mentions) Cacl2 2h2o, by Merck Group (1 mentions)

Spelling variants of this product

Matrigel (ECM gel, (9 citations) ECM Gel from Engelbreth-Holm-Swarm murine sarcoma (7 citations) ECM Matrix gel solution (6 citations) ECM gel from Engelbreth-Holm-Swarm mouse sarcoma (2 citations) ECM gel (E1270) (2 citations) ECM Gel-coated (2 citations) ECM gel drop (1 citations)

88 protocols using ecm gel

Aortic Ring Angiogenesis Assay

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Aortas from 12-days old chicken embryos were harvested under sterile conditions and cut into 4–5 rings of <1 mm length after removing fat and membranous tissue from around the vessel (~4–5 mm long). Rings were embedded in 80 μL of extracellular matrix gel (ECM gel, Sigma Aldrich) in wells of 48 well plates. After allowing the ECM gel to gel at 37°C for 20 min, alpha MEM media (Sigma Aldrich) supplemented with 2% FCS (Sigma Aldrich) was gently added to the well.
The dissolution extracts of MBG and Cu-MBG were prepared by suspending the powder at different concentrations (1 mg/mL, 300 μg/mL, 100 μg/mL, and 30 μg/mL) in alpha MEM growth media at 4°C for 72 h. After the incubation the powders were removed from the suspension by filtration, and the resulting solution extracts were used for the aortic ring experiments. VEGF was used as a positive control to treat aortic rings at 3, 10, and 30 ng/mL dosage.
Outgrowth from rings was captured by photography (40x magnification) every 24 h for 5 days and 48 h was chosen as a time point to compare outgrowth between the tested particle extracts and controls. Outgrowth was measured by tracing the outline of the area covered by the outgrowth of endothelial sprouts/network then subtracting the area covered by the original aortic ring using ImageJ software.

Paterson T.E., Bari A., Bullock A.J., Turner R., Montalbano G., Fiorilli S., Vitale-Brovarone C., MacNeil S, & Shepherd J. (2020). Multifunctional Copper-Containing Mesoporous Glass Nanoparticles as Antibacterial and Proangiogenic Agents for Chronic Wounds. Frontiers in Bioengineering and Biotechnology, 8, 246.

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Polyphosphate Effects on Keratinocytes and Endothelial Cells

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Polyphosphate (polyP; sodium salt; food grade, purity ≥ 99%,) with an average chain length of 40 phosphate units was purchased from Chemische Fabrik Budenheim (Budenheim, Germany). Na₂CO₃ (#222321), CaCl₂·2H₂O (#223506), fetal bovine serum (FBS; #F7942), human epidermal keratinocytes (HEK; #102-05A), human epidermal keratinocyte complete culture medium (#SCMK001), EpiGR human epidermal keratinocyte complete culture media kit (#SCMK001), ECM gel (#E6909; ECM gel from Engelbreth-Holm-Swarm murine sarcoma), MTT (thiazolyl blue tetrazolium bromide; #M2128), and 96-well plates (#CLS3595) were from Sigma-Aldrich (Taufkirchen, Germany). Cultrex basal membrane extract (BME; growth factor reduced) was from R&D systems (Abingdon; UK). Human umbilical vein endothelial cells (HUVEC) and bovine brain extract (#CC-4098) were from Lonza (Köln, Germany).

Wang S., Neufurth M., Schepler H., Tan R., She Z., Al-Nawas B., Wang X., Schröder H.C, & Müller W.E. (2023). Acceleration of Wound Healing through Amorphous Calcium Carbonate, Stabilized with High-Energy Polyphosphate. Pharmaceutics, 15(2), 494.

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Endothelial Progenitor Cell Tube Formation

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An ECM gel (Sigma) placed on a 96-well culture plate at 37 °C for 1 h to allow solidification after thawed at 4 °C overnight. EPCs treated with or without PA were harvested and seeded (10,000 cells/well) on the top of the solidified ECM gel in EBM-2 medium supplemented with 0.5% BSA and VEGF (100 ng/ mL). Cells were incubated at 37 °C for 12 h. Tube formation was defined as a structure exhibiting a length 4 fold more than its width. The networks of tubes were photographed from six randomly chosen fields with a microscope. The total length of the tube structures in each photograph was measured using Adobe Photoshop software (Adobe, San Jose, CA).

Guo W., Li Z., Xie X., Qin T., Wu Y., Li Z., Chai J., Yi F., Tan T., Zhu H, & Wang S. (2015). Urinary Trypsin Inhibitor Attenuates Acute Lung Injury by Improving Endothelial Progenitor Cells Functions. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, 36(3).

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Cell Migration Assay with Transwell

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Transwell chambers (8 μm pores, Corning, USA) were pre-coated with the ECM Gel (E1270, Sigma-Aldrich, USA). 2 × 10⁴ cells were seeded in the upper chambers with serum-free medium, and normal medium in the bottom chambers. After 12 h of incubation, cells adherent to the lower surface of the chambers were stained with 0.4% crystal violet for photographing and quantitative analysis.

Lü J., Zhao Q., Guo Y., Li D., Xie H., Liu C., Hu X., Liu S., Hou Z., wei X., Zheng D., Pestell R.G, & Yu Z. (2023). Regulation of ERα-dependent breast cancer metastasis by a miR-29a signaling. Journal of Experimental & Clinical Cancer Research : CR, 42, 93.

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In Vitro Angiogenesis and Migration Assays

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HUVECs were seeded in a 96-well plate coated with cold ECM gel (Sigma-Aldrich). Thereafter, the HUVECs were treated with the indicated supplements. Tube formation assay was performed, and the results were examined under a microscope (Leica, Germany) after 12 hours.
HUVECs (3 × 10⁴/well) were cultured in 200 μl basal medium containing 1% FBS. The cells were then seeded into the upper chamber of a transwell plate (Corning, China). The lower chamber was supplemented with basal medium containing 5% FBS and the indicated treatments. After 24 hours, the upper chamber was fixed with 4% paraformaldehyde for 30 minutes, washed with PBS three times, and stained with 0.5% crystal violet (Beyotime, China) for two minutes. Cell migration in the transwell assay was examined using an optical microscope.
HUVECs were cultured in a six-well plate until they reached confluence. A 200 μl sterile loading gun was used to divide the cells to form a cell-free area in the centre of the culture plate. PBS was used to wash the cellular debris three times. Thereafter, the cells were cultured in basal medium containing 1% FBS and the indicated treatment. The scratch wound assay was performed immediately thereafter and 12 hours later, and the results were observed using a microscope.

Zhu D., Fang H., Yu H., Liu P., Yang Q., Luo P., Zhang C., Gao Y, & Chen Y.X. (2022). Alcohol-induced inhibition of bone formation and neovascularization contributes to the failure of fracture healing via the miR-19a-3p/FOXF2 axis. Bone & Joint Research, 11(6), 386-397.

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Metabolic Profiling of Human Islets

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The oxygen consumption rates (OCRs) and extracellular acidification rates (ECARs) of isolated human islets were determined using the Extracellular Flux Analyzer xFe96 (Seahorse Bioscience, Billerica, MA, USA). Human islets were precultured without or with GDF15 (10 ng/mL or 100 ng/mL) for 24 h and then with an additional mixture of cytokines (IL-1β 20 ng/mL + IFN-γ 20 ng/mL) for 48 h. After the treatment, 8–10 human islets (6 replicates for each treatment group) were placed into each well of the xFe 96 cell culture microplate precoated with 3% ECM gel (Sigma-Aldrich) and 5 μg/mL fibronectin (Sigma-Aldrich). Islets were preincubated for 2 h at 37 °C in Seahorse Biosciences’ XF assay medium supplemented with 5.5 mM glucose. Following 20 min of basal recording, the cells were sequentially injected with oligomycin (Sigma Aldrich), carbonyl cyanide-p-trifluoromethoxyphenyl-hydrazone (FCCP) (Sigma Aldrich), and a combination of rotenone (Sigma Aldrich) and antimycin A (Sigma Aldrich) at intervals of 20 to 30 min. Each substance was introduced at the concentration of 5 µM. Measurements of the ATP-coupled OCR, maximum respiratory capacity, and ECAR were used to assess mitochondrial function. The data were normalized to protein contents determined using the DC protein assay (Bio-Rad).

Ngamjariyawat A., Cen J., Wang X, & Welsh N. (2024). GDF15 Protects Insulin-Producing Beta Cells against Pro-Inflammatory Cytokines and Metabolic Stress via Increased Deamination of Intracellular Adenosine. International Journal of Molecular Sciences, 25(2), 801.

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Differential Plating for Spermatogonial Stem Cell Isolation

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Six-well plates were coated with extracellular matrix (ECM) gel (Sigma-Aldrich, USA) as the manufacturer indicated. The ECM gel was prepared from Engelbreth-Holm-Swarm (EHS) mouse sarcoma and was composed of laminin as the major component, collagen type IV, heparan sulfate proteoglycan, entactin and other minor components. Cells were seeded at concentrations of 1,000,000 cells per well containing DMEM with antibiotics and 10% KSR. The plates were incubated at 37°C in a humidified atmosphere with 5% CO₂. In the germ cell-removed group, the SSCs were removed from the in vitro culture using the differential plating method described by He et al, in which following 3 hr of incubation, somatic cells attached to the plate but SSCs had remained in the suspension and consequently were eliminated from the plate 26 (link). No intervention was performed in the control group. To further evaluate the effectiveness of the differential plating method for removal of SSCs, the gene expression of spermatogonial markers was assessed. In this context, Thy1 and Bcl6b were used as the markers of undifferentiated spermatogonia 27 (link) and ckit was used as a marker for differentiated spermatogonia 28 (link). The medium was replaced with a fresh one every three days in both groups.

Akbarinejad V., Tajik P., Movahedin M, & Youssefi R. (2016). Effect of Removal of Spermatogonial Stem Cells (SSCs) from In Vitro Culture on Gene Expression of Niche Factors in Bovine. Avicenna Journal of Medical Biotechnology, 8(3), 133-138.

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Kidney Spheroid Formation and Assays

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Immortalized renal cells were suspended in a 2:1 mixture of ECM gel (Sigma) and matrigel and 50 μL droplets at 1~2 × 10⁴ cells/μL were deposited on the inverted lid of a culture dish. Then the lid was placed onto a phosphate-buffered saline (PBS)-filled dish, and after gelation (3–6 h later), cell in the matrix were transferred to the SFU filled with DMEM/F12 supplemented with 10% FBS, 1X Insulin-Transferrin-Selenium (ITS), 20 ng/mL epidermal growth factor (EGF), 100 nM dexamethasone, 20 uM 1α,25-Dihydroxyvitamin D3 and 5 μM all-trans retinoic acid (ATRA) for 6 h. Cells were then cultured for 2–5 additional days without ATRA. For nephrotoxicity tests, kidney spheroids were treated with various concentrations of drugs in serum-free DMEM/F12 for 24 h, and for transporter activity assays, spheroids were pretreated overnight with cimetidine or verapamil before drug treatment.

Kang H.M., Lim J.H., Noh K.H., Park D., Cho H.S., Susztak K, & Jung C.R. (2019). Effective reconstruction of functional organotypic kidney spheroid for in vitro nephrotoxicity studies. Scientific Reports, 9, 17610.

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Integrin Signaling in PSMA-Knockout Cells

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For analysis of signaling responses affected by the ligation of integrins in 22RV1-CRISPR-PSMA^knockout adherent cells, the cells and scramble control were washed with PBS and starved from serum for 4 hours before being stimulated with culture medium containing 10% FBS. For analysis of signaling responses affected by the ligation of integrins in nonadherent cells, confluent 22RV1-CRISPR-PSMA^knockout cells and scramble control were detached with trypsin and then washed with PBS. Cells were resuspended in serum-free medium containing 5% ECM Gel (Sigma-Aldrich) and maintained in suspension for 4 hours before stimulation with culture medium containing 10% FBS.

Caromile L.A., Dortche K., Rahman M.M., Grant C.L., Stoddard C., Ferrer F.A, & Shapiro L.H. (2017). PSMA redirects cell survival signaling from the MAPK to the PI3K-AKT pathways to promote the progression of prostate cancer. Science signaling, 10(470), eaag3326.

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Cell Invasion Assay Protocol

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After 48 h of transfection, the cells (0.1X10⁶) were washed with PBS and resuspended in serum-free DMEM. Cells were then added onto the precoated upper chamber (with 2 mg/ml ECM gel; Sigma-Aldrich, Missouri, USA) of transwell plate (Corning, New York, USA), and in lower chamber DMEM supplemented with 10% FBS was added. Cells were allowed to invade for 48 h in CO₂ incubator. Cells that remained in the upper chamber were removed whereas the cells which invaded through the membrane were first fixed with 5% glutaraldehyde and then stained with 0.2% crystal violet solution in 2% ethanol. The cells were then counted under the phase contrast inverted microscope for quantification.

Singh G., Sharma S.K., Dorata A, & Singh S.K. (2023). miR-17 ~ 92 suppresses proliferation and invasion of cervical cancer cells by inhibiting cell cycle regulator Cdt2. Discover. Oncology, 14, 172.

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