Matrigel fibronectin

Manufactured by Merck Group

Sourced in France

Matrigel-fibronectin is a protein mixture that supports the attachment and growth of various cell types. It is commonly used as a component in cell culture and tissue engineering applications. The product provides a natural extracellular matrix environment to facilitate cell attachment, proliferation, and differentiation.

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Lab products found in correlation

Transferrin, by Merck Group (2 mentions) 2 mercaptoethanol, by Bio-Rad (2 mentions) Bsa fraction 5, by Roche (2 mentions) Sodium selenite, by Merck Group (2 mentions) Penicillin, by Lonza (1 mentions) Streptomycin, by Lonza (1 mentions) Hek293 cells crl 1573, by American Type Culture Collection (1 mentions) Plasmocin prophylactic, by InvivoGen (1 mentions) Fetal bovine serum (fbs), by Bio-Rad (1 mentions) Mycoalert mycoplasma detection kit, by Lonza (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Nicotinamide, by Merck Group (1 mentions) Penicillin, by GE Healthcare (1 mentions) Streptomycin, by GE Healthcare (1 mentions)

Spelling variants of this product

Fibronectin (1886 citations) Matrigel (1099 citations) Matrigel/fibronectin-coated (3 citations)

4 protocols using matrigel fibronectin

Culturing EndoC-βH1 and HEK293 cells

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The EndoC‐βH1 human pancreatic β cell line was purchased from Human Cell Design (https://www.humancelldesign.com). Cells were cultured and split as previously described.^[⁷⁶^] Briefly, EndoC‐βH1 cells were seeded at an approximate density of 70,000‐75,000 cells cm⁻² on culture plates pre‐coated with Matrigel‐fibronectin (100 mg L⁻¹ and 2 mg mL⁻¹, respectively, Sigma–Aldrich) at 37 °C, 5% CO₂ in complete OPTIβ1 medium (Human Cell Design). Cells were passed every 7 days. For transfection, DMEM medium was used containing 2% FBS, 5.6 mmol L⁻¹ glucose, 50 µmol L⁻¹ 2‐mercaptoethanol (Biorad, CA, USA), 10 mmol L⁻¹ nicotinamide (Calbiochem, Darmstadt, Germany), 5.5 µg mL⁻¹ transferrin, 6.7 ng mL⁻¹ selenite (Sigma–Aldrich).
HEK293 cells (CRL‐1573) were purchased from the American Type Culture Collection (ATCC; https://www.atcc.org). Cells were cultured in DMEM supplemented with 10% FBS and 100 units mL⁻¹ penicillin and 100 µg mL⁻¹ streptomycin (Lonza). The same medium without antibiotics was used for transfection.

González‐Moro I., Garcia‐Etxebarria K., Mendoza L.M., Fernández‐Jiménez N., Mentxaka J., Olazagoitia‐Garmendia A., Arroyo M.N., Sawatani T., Moreno‐Castro C., Vinci C., Op de Beek A., Cnop M., Igoillo‐Esteve M, & Santin I. (2023). LncRNA ARGI Contributes to Virus‐Induced Pancreatic β Cell Inflammation Through Transcriptional Activation of IFN‐Stimulated Genes. Advanced Science, 10(25), 2300063.

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Pancreatic Cell Culture and Islet Isolation

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The EndoC-βH1 human pancreatic cell line (Univercell Biosolutions, Paris, France) was cultured in plates coated with Matrigel-fibronectin (100 mg/ml and 2 mg/ml, respectively; Sigma-Aldrich, Burlington, USA) in Opti-β1 medium (Univercell Biosolutions). DMEM containing 5.6 mmol/l glucose, 2% vol/vol Fetal Bovine Serum, 50 μmol/l 2-mercaptoethanol (Bio-Rad, Hercules, USA), 10 mmol/l nicotinamide (Calbiochem, Darmstadt, Germany), 5.5 μg/ml transferrin and 6.7 ng/ml selenite (Sigma-Aldrich) was used for transfection.
EndoC-βH1 cell line was Mycoplasma free as determined by the MycoAlert Mycoplasma Detection kit (Lonza). For the prevention of Mycoplasma contamination, Plasmocin Prophylactic (Invivogen, Toulouse, France) was added to the culture medium on a regular basis.
cDNA samples from human pancreatic islets were obtained from Cisanello University Hospital, Pisa, Italy. All the islets were isolated and cultured using the same experimental conditions and following established isolation procedures (15 (link)). Characteristics of islet preparations are described in Table S1. The Ethical Committee of Cisanello University Hospital approved experiments using human islets.

González-Moro I., Rojas-Márquez H., Sebastian-delaCruz M., Mentxaka-Salgado J., Olazagoitia-Garmendia A., Mendoza L.M., Lluch A., Fantuzzi F., Lambert C., Ares Blanco J., Marselli L., Marchetti P., Cnop M., Delgado E., Fernández-Real J.M., Ortega F.J., Castellanos-Rubio A, & Santin I. (2023). A long non-coding RNA that harbors a SNP associated with type 2 diabetes regulates the expression of TGM2 gene in pancreatic beta cells. Frontiers in Endocrinology, 14, 1101934.

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Culture of EndoC-βH1 Insulin-Secreting Cells

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EndoC-βH1 cells (EndoC-βH1 cells, Paris, France)⁸⁸ (link) were grown in Matrigel fibronectin-coated (100 μg/mL and 2 μg/mL, respectively, Sigma–Aldrich, Steinheim, Germany) culture vessels in DMEM (Thermo Fisher Scientific, Waltham, MA, USA) containing 5.6 mM glucose, 2% BSA fraction V (Roche Diagnostics, Mannheim, Germany), 10 mM nicotinamide (Merck Millipore, Darmstadt, Germany), 50 μM 2-mercaptoethanol, 5.5 μg/mL transferrin, 6.7 ng/mL sodium selenite (Sigma–Aldrich), 100 U/mL penicillin, and 100 μg/mL streptomycin (PAA Laboratories, Pasching, Austria). The cells were incubated in a humidified atmosphere with 5% CO² at 37°C. The cells were tested for mycoplasma contamination on a regular basis.

Karagiannopoulos A., Westholm E., Ofori J.K., Cowan E., Esguerra J.L, & Eliasson L. (2023). Glucocorticoid-mediated induction of ZBTB16 affects insulin secretion in human islets and EndoC-βH1 β-cells. iScience, 26(5), 106555.

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MICU2 Knockdown in Beta Cell Lines

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INS-1 832/13 cells were cultured as previously described [20 (link)]. EndoC-βH1 cells [21 ] were grown on Matrigel-fibronectin-coated (100 μg/mL and 2 μg/mL, respectively, Sigma–Aldrich) culture vessels in DMEM containing 5.6 mM of glucose, 2% BSA fraction V (Roche Diagnostics), 10 mM of nicotinamide (Calbiochem), 50 μM of 2-mercaptoethanol, 5.5 μg/mL of transferrin, 6.7 ng/mL of sodium selenite (Sigma–Aldrich), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (PAA). Both cell lines were cultured at 37 °C with 5% CO₂. Unless otherwise stated, EndoC-βH1 cells were seeded at 2.3 × 10⁵ cells/cm² and INS-1 832/13 cells at 1.5 × 10⁵ cells/cm² in 24-well plates (Matrigel-fibronectin coated or uncoated) and transfected either with siRNA specific to MICU2 (50 nM) or a scrambled siRNA (50 nM). Forty-eight h after knockdown, the growth media were changed to an overnight starvation medium containing 5.6 mM of glucose for INS-1 832/13 cells and 1 mM of glucose for EndoC-βH1 cells. HEK-293T cells were cultured in DMEM containing 1 g/L of d-glucose and supplemented with 10% heat-inactivated FBS, 100 U/mL of penicillin, and 100 μg/mL of streptomycin (PAA) [22 ]. The cells were plated at 1.5 × 10⁵ cells/cm² in 24-well plates and transfected with either siRNA specific to MICU2 or a scrambled siRNA. All of the assays were performed 48–72 h after knockdown.

Vishnu N., Hamilton A., Bagge A., Wernersson A., Cowan E., Barnard H., Sancak Y., Kamer K.J., Spégel P., Fex M., Tengholm A., Mootha V.K., Nicholls D.G, & Mulder H. (2021). Mitochondrial clearance of calcium facilitated by MICU2 controls insulin secretion. Molecular Metabolism, 51, 101239.

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