Matrigel coated

Manufactured by Corning

Sourced in United States, Canada, China

Matrigel-coated lab equipment is a specialized surface coating used to support the growth and attachment of cells in cell culture applications. It provides an extracellular matrix-like environment for cells to adhere to and proliferate. The coating is made from a gelatinous protein mixture secreted by Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells.

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Matrigel (8653 citations) Matrigel-coated Transwell inserts (36 citations) Matrigel-coated 96-well plates (14 citations) Matrigel matrix-coated (6 citations) Matrigel-coated Boyden chambers (5 citations) Matrigel pre-coated transwell inserts (4 citations) Matrigel-coated tissue culture plates (3 citations) Matrigel-coated Transwell invasion assay plates (2 citations) Matrigel-coated nucleopore filter inserts in a 24-well transwell chamber (2 citations) Matrigel-coated 24-well Transwell plates (2 citations)

141 protocols using matrigel coated

Comprehensive iPSC Characterization Protocol

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iPSC pluripotency was assessed by staining iPSC colonies cultured in 6-well plates for alkaline phosphatase using the Vector Blue Alkaline Phosphatase Kit (Reactorlab, #SK‒5300), 4 – 6 days after splitting. iPSCs seeded into Matrigel-coated (Corning, #356230) 8-well chamber slides (ibidi, #80826) were further stained for the pluripotency markers SOX2, OCT4, NANOG and SSEA4 (additional antibody information is in the table ‘Key Resources Table’). The potential to differentiate into the three embryonic germ layers was assessed by directed differentiation into ectoderm, mesoderm and endoderm using the STEMdiff Trilineage Differentiation Kit (STEMCELL Technologies, #05230) according to the manufacturer’s instructions on cells maintained in Matrigel-coated (Corning, #356230) 8-well chamber slides (ibidi, #80826). To test for chromosomal aberrations, G-banded karyotyping was performed on 20 cells per line (Cell Guidance Systems, UK). Further, array comparative genomic hybridization (array CGH) was performed using the Illumina HumanOmni2.5Exome-8 BeadChip v1.3 (LIFE & BRAIN GmbH, Germany) on genomic DNA extracted from iPSCs by the DNeasy Blood & Tissue Kit (QIAGEN, #69504).

Cowan C.S., Renner M., De Gennaro M., Gross-Scherf B., Goldblum D., Hou Y., Munz M., Rodrigues T.M., Krol J., Szikra T., Cuttat R., Waldt A., Papasaikas P., Diggelmann R., Patino-Alvarez C.P., Galliker P., Spirig S.E., Pavlinic D., Gerber-Hollbach N., Schuierer S., Srdanovic A., Balogh M., Panero R., Kusnyerik A., Szabo A., Stadler M.B., Orgül S., Picelli S., Hasler P.W., Hierlemann A., Scholl H.P., Roma G., Nigsch F, & Roska B. (2020). Cell Types of the Human Retina and Its Organoids at Single-Cell Resolution. Cell, 182(6), 1623-1640.e34.

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Cell Invasion and Migration Assay

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The invasive and migration abilities of the cells were analyzed by Transwell chamber assays with or without coated matrigel (Corning). 2.5 × 10⁴ TPC1 cells and 2 × 10⁴ HTori-3 cells were harvested 3 days after transfection with a solution of PBS/5 mM EDTA/5 mM EGTA. 22-hours after seeding into the upper chamber, passing cells were fixed, stained by using Diff-Quick stain kit (Polysciences) and quantified by counting and averaging 5 separate spots of 4 mm diameter.

Augenlicht A., Saiselet M., Decaussin-Petrucci M., Andry G., Dumont J.E, & Maenhaut C. (2021). MiR-7-5p inhibits thyroid cell proliferation by targeting the EGFR/MAPK and IRS2/PI3K signaling pathways. Oncotarget, 12(16), 1587-1599.

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Transwell Invasion Assay for Cell Migration

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The transwell invasion assay was performed in 24-well plates with a 6.5mm insert transwell chamber with 8μm polycarbonate membrane (Corning) pre-coated Matrigel (Corning). The single cell suspension was added into to upper chamber with 5*10⁴ cells in 200μl culture medium with 2% FBS, and 500μl culture medium with 20% FBS was added into the lower chamber. After 48 h, discarded the solution in the upper chamber and wiped the upper layer of the membrane. Move the chamber into 4% PFA to fix for 5 min. Stain the membrane with crystal violet for 5 min. Finally, obtained the photographs on the microscope. All experiments were performed in triplicate.

Yu M., Chang Y., Zhai Y., Pang B., Wang P., Li G., Jiang T, & Zeng F. (2023). TREM2 is associated with tumor immunity and implies poor prognosis in glioma. Frontiers in Immunology, 13, 1089266.

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Cell Migration and Invasion Assay

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Cell migration and invasion assays were performed in 24-well plates with 8 μm pore size chamber inserts (Millipore), as previously described [27 (link)]. The invasion assay used coated Matrigel (Corning, 1:10 dilution) in the upper chamber, simulating characteristics of the extracellular matrix, and the migration assay used chamber inserts only. MDA-MB-231 cells were transfected with LINC00449 and LINC01270 siRNAs and NC siRNA for 16 h. Then, siRNA-transfected cells were reseeded into the upper chamber with 100 μL serum-free medium (6 × 104 cells/well). A total of 750 μL medium containing 10% FBS was placed in the lower chamber as a chemoattractant. After incubation at 37 °C for 48 h, cells adhering to the lower surface membrane of the upper chamber were fixed in 10% neutral buffered formalin for 30 min, followed by staining with 0.1% crystal violet (Sigma-Aldrich), and then subjected to microscopic inspection. Five visual fields of each insert were randomly chosen. crystal violet was then eluted using 33% acetic acid and quantified by measuring the absorbance at 590 nm with BioTek Synergy HT plate reader. Relative migrated and invaded cells were normalized to the NC siRNA group and expressed as percent (%) of negative control. Results represent data obtained from three independent experiments.

Ping J., Huang S., Wu J., Bao P., Su T., Gu K., Cai H., Guo X., Lipwort L., Blot W.J., Zheng W., Cai Q, & Shu X.O. (2020). Association between lincRNA expression and overall survival for patients with triple-negative breast cancer. Breast cancer research and treatment, 186(3), 769-777.

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3D Retinal Tissue Generation from hiPSCs

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HiPSC cell line: SB line (Cellapy, CA4002106). The cells were maintained in mTeSR™1 medium (STEMCELL Technologies 05850) at 37°C in the presence of 5% CO₂ and were induced to differentiate into three-dimensional retinal tissue following previous protocols [11 (link)]. Briefly, iPS cells were induced to form embryoid bodies (EBs) and sequentially differentiated in N2 (Gibco, 17502-048) and B27 (Gibco, 12587-010) supplements (Figure 1(b)).
The PLGA scaffolds used in the present study were prepared by electrospinning and hydrophilic treatment (Figure 1(c)). The molecular weight was designed as 35,000 to meet the requirement of complete degradation in two months. The pores were formed by nanofibers (Figure 1(d)), providing space for cell adherence and migration. Subsequently, three-dimensional retinal tissue with retinal progenitors was seeded and adhered onto Matrigel-coated (Corning, 354277) PLGA scaffolds (approximately 2 mm × 4 mm × 0.1 mm). Then, the progenitors and PLGA composite were maintained in retinal differentiation medium (supplemented with B27 and 10% FBS (Gibco, 10099-141)) for another five days under 37°C in the presence of 5% CO₂ until transplantation (Figure 1(e)).

Luo Z., Li K., Li K., Xian B., Liu Y., Yang S., Xu C., Fan Z., Lu S., Zhang H, & Ge J. (2018). Establishing a Surgical Procedure for Rhesus Epiretinal Scaffold Implantation with HiPSC-Derived Retinal Progenitors. Stem Cells International, 2018, 9437041.

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Culturing Pluripotent Stem Cells in E8 Medium

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Cells were maintained in home-made E8 medium (DMEM/F12, L-ascorbic acid, selenium, transferrin, insulin, fibroblast growth factor 2 [FGF2], and TGFβ) following published protocols (Chen et al., 2011 (link)). In brief, cells were cultured in E8 medium (with 1× penicillin/streptomycin) on Matrigel-coated (Corning, 354230) plates. Confluent (∼80%) cells were passaged every 3–4 days using the DPBS-EDTA method as described by Beers et al. (2012) (link). ROCK inhibitor Y-27632 (5 μM) was used during cell passaging.
Unless otherwise stated, all the pyruvate concentrations in the article refer to the amount of pyruvate added to the medium (surplus to the 0.5 mM pyruvate in E8 medium).

Song C., Xu F., Ren Z., Zhang Y., Meng Y., Yang Y., Lingadahalli S., Cheung E., Li G., Liu W., Wan J., Zhao Y, & Chen G. (2019). Elevated Exogenous Pyruvate Potentiates Mesodermal Differentiation through Metabolic Modulation and AMPK/mTOR Pathway in Human Embryonic Stem Cells. Stem Cell Reports, 13(2), 338-351.

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Generation of ABCA3 Mutant iPSCs

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All human iPSCs were maintained in feeder-free conditions, cultured on Matrigel-coated (Corning) plates in mTeSR media (StemCell Technologies), and passaged using Gentle Cell Dissociation Reagent (StemCell Technologies). Reprogramming of the BU3 human iPSC line was previously reported in Kurmann et al. (65 (link)), and editing of this line to target an ABCA3:GFP fusion cassette to the endogenous ABCA3 locus (BU3^ABCA3:GFP) was previously reported in Sun et al. (39 (link)). For derivation of ABCA3 mutant patient-specific iPSC lines, dermal fibroblasts from each individual were received from Washington University School of Medicine. Genetic evaluation found no mutations in other genes associated with surfactant production, such as SFTPC or SFTPB genes. Reprogramming of dermal fibroblasts from individuals with homozygous E690K or homozygous W308R ABAC3 mutations was performed as detailed in the Supplemental Methods.

Sun Y.L., Hennessey E.E., Heins H., Yang P., Villacorta-Martin C., Kwan J., Gopalan K., James M., Emili A., Cole F.S., Wambach J.A, & Kotton D.N. (2024). Human pluripotent stem cell modeling of alveolar type 2 cell dysfunction caused by ABCA3 mutations. The Journal of Clinical Investigation, 134(2), e164274.

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Cultivation of SiMa and IMR90-4 Cells

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SiMa cells [17 (link)] were cultivated in RPMI1640 GLutaMAX™ medium (Gibco, Grand Island, NY, USA) with 1% penicillin/streptomycin (P/S, PAN Biotech, Aidenbach, Germany), and 10% Fetal Bovine Serum (FBS, PAN Biotech, Aidenbach, Germany) on untreated cell culture plates. Medium was changed every 2–3 days and upon reaching 80–90% confluency, cells were split 1:15 after trypsinization (Trypsin-EDTA, Gibco, Grand Island, NY, USA). Undifferentiated IMR90-4 cells [18 (link)] were cultured on Matrigel-coated (Corning, NY, USA) plates in StemMACs™ iPS-Brew XF human cell culture medium (Miltenyi, Bergisch Gladbach, Germany) containing 1% P/S in feeder-free conditions. Medium was changed every 2 days and cells were split 1:40 upon reaching 80–90% confluency with 0.5 mM EDTA (Sigma-Aldrich, Darmstadt, Germany) in PBS. Newly split cells were supplemented with 1 µM Y-27632 ROCK inhibitor (Bertin Pharma, Montigny le Bretonneux, France). Matrigel was applied to the appropriate cell-culture plates (96/24/6-well plates, Sarstedt, Nümbrecht, Germany) and incubated at room temperature for at least 2 h before use. The Matrigel solution was prepared on ice by diluting previously prepared aliquots 1:100 with KnockOut Medium (Thermo Fisher Scientific, Waltham, MA, USA).

Schjeide B.M., Schenke M., Seeger B, & Püschel G.P. (2022). Validation of a Novel Double Control Quantitative Copy Number PCR Method to Quantify Off-Target Transgene Integration after CRISPR-Induced DNA Modification. Methods and Protocols, 5(3), 43.

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Isolation and Differentiation of Myoblasts

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were isolated from wild-type and LAKI-Nanog (LmnaG609G/G609G, Col1a1 tetO-Nanog/+; ROSA26rtTA/rtTA) mice as described previously^{61 (link)}. Isolated myoblast cells were then plated onto matrigel-coated (0.1 mg/ml) (CORNING, Corning, NY) dishes. High glucose Dulbecco’s Modified Eagle Medium (DMEM) was used for myoblast culture (Waltham, MA), 20% fetal bovine serum (FBS, Atlanta Biologicals, Flowery Branch, GA), 10% horse serum (Grand Island, NY), 0.5% chicken embryo extract (CEE, Accurate Chemical and Scientific, Westbury, NY), 2.5 ng/ml bFGF (ORF Genetics, Iceland), 10 μg/ml gentamycin (Waltham, MA), and 1% Antibiotic-Antimycotic (Grand Island, NY), and 2.5 μg/ml plasmocin prophylactic (Invivogen, San Diego, CA).
For differentiation, the cells were allowed to reach 100% confluence (in 2–3 days) and then the medium was switched to differentiation medium that was composed of DMEM with high glucose, 5% horse serum (HS), and 1% AA to promote the formation of multinucleated myotubes.

Rajabian N., Ikhapoh I., Shahini S., Choudhury D., Thiyagarajan R., Shahini A., Kulczyk J., Breed K., Saha S., Mohamed M.A., Udin S.B., Stablewski A., Seldeen K., Troen B.R., Personius K, & Andreadis S.T. (2023). Methionine adenosyltransferase2A inhibition restores metabolism to improve regenerative capacity and strength of aged skeletal muscle. Nature Communications, 14, 886.

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Pluripotent Stem Cell Culture Protocols

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Human iPSCs were cultured on Matrigel-coated (Corning) 6-well plates in FTDA culture medium at 5% CO2, 5% O2 as described in^{34 (link)–37 (link)}. Mouse iPSCs were cultured either in feeder-dependent conditions or in feeder-free conditions (2i)³⁸. For feeder-dependent conditions (ES feeder medium) Knockout^TM DMEM (KO-DMEM; Life-Technologies) was supplemented with 15% FBS, 1% P/S, 1% GlutaMAX, 1% NEAA, 1% Sodium Pyruvate, 1% β-Mercaptoethanol and 240 U/ml leukaemia inhibitory factor (LIF, Cell guidance systems). In case of feeder-free (2i) culture mouse KO-DMEM with 15% Knockout Serum Replacement (KOSR, Life Technologies), 1% P/S, 1% GlutaMAX, 1% NEAA, 1% Sodium Pyruvate, 1% β-Mercaptoethanol, 240U/ml LIF and 1 μM PD0325901 (Calbiochem) and 3 μM GSK3β-inhibitor CHIR99021 (Axon Medchem) was used.

Raab S., Klingenstein M., Möller A., Illing A., Tosic J., Breunig M., Kuales G., Linta L., Seufferlein T., Arnold S.J., Kleger A, & Liebau S. (2017). Reprogramming to pluripotency does not require transition through a primitive streak-like state. Scientific Reports, 7, 16543.

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