Matrigel

Manufactured by Selleck Chemicals

Sourced in United States

Matrigel is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma. It is used as a substrate for the attachment, growth, and differentiation of cells in in vitro cell culture applications.

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Lab products found in correlation

Matrigel, by BD (3 mentions) Matrigel, by Corning (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Accutase, by STEMCELL (2 mentions) Accutase, by Thermo Fisher Scientific (1 mentions) Chir99021, by Selleck Chemicals (1 mentions) Mtesr1, by STEMCELL (1 mentions) Mtesr1, by Selleck Chemicals (1 mentions) Advanced dmem f12, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Neurobasal, by Thermo Fisher Scientific (1 mentions) Compound c, by Selleck Chemicals (1 mentions) Retinoic acid, by Selleck Chemicals (1 mentions) Transit lt1 transfection reagent, by Mirus Bio (1 mentions) Y 27632, by Selleck Chemicals (1 mentions) Mtesr media, by STEMCELL (1 mentions) Mitomycin c, by Selleck Chemicals (1 mentions) Giemsa, by Solarbio (1 mentions) Rock inhibitor y 27632, by Selleck Chemicals (1 mentions)

5 protocols using matrigel

Cardiac Progenitor Differentiation from hPSCs

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Human LQTS iPSCs (iPSCORE_2_1^{45 (link)}) and human ESCs (H1 and H9^{46 (link)}) were maintained on Matrigel (Corning)-coated surfaces in mTeSR1 (STEMCELL Technologies) as previously decribed^{47 (link)}. Cardiac progenitor differentiation was performed as described previously²². Briefly, hPSCs were singularized with Accutase (Thermo Fisher Scientific) and plated onto Matrigel-coated plates at a density of 3.0×10⁵ cells/cm² in mTeSR1 supplemented with 10 μM Rho-associated protein kinase (ROCK) inhibitor Y-27632 (Selleckchem) on day −2. The differentiation was initiated with Wnt activation by 8 μM CHIR99021 (Selleckchem) on day 0 and sequential Wnt inhibition by 5μM IPW2 on day 3. Cardiac progenitors were ready on day 6 for CS generation.

Qian T., Gil D.A., Guzman E.C., Gastfriend B.D., Tweed K.E., Palecek S.P, & Skala M.C. (2020). Adaptable pulsatile flow generated from stem cell-derived cardiomyocytes using quantitative imaging-based signal transduction. Lab on a chip, 20(20), 3744-3756.

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Differentiation of H9 ESCs to NPSCs

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H9-ESCs were induced to differentiate into H9–NPSCs as previous reported [23 (link)]. Briefly, H9-ESCs cultured in E8 medium were passaged in a six well plate coated with Matrigel (BD Biosciences, USA). H9-ESCs at about 30% confluency were cultured in basic medium containing 1 μM compound C (Selleck, USA). After three days, 100 nM retinoic acid was added to the basic medium. After another three days, 100 ng/mL SHH and 100 nM retinoic acid were both added to the basic medium without compound C. After another three days of differentiation, H9-ESCs had almost differentiated into H9–NSPCs. Basic medium was composed of 50% Advanced DMEM/F12 (Invitrogen, USA), 50% Neurobasal (Gibco, USA), 1 × N2 (Gibco, USA), 1 × B27 (Gibco, USA), 1% GlutaMAX (Gibco, USA), 1% NEAA (Gibco, USA), 1% penicillin-streptomycin (Gibco, USA). H9–NSPCs were then cultured on plates coated with Matrigel in basic medium containing 100 nM retinoic acid (Selleck, USA) and 0.5 μM small purmorphamine (Selleck, USA).

Xu B., Yin M., Yang Y., Zou Y., Liu W., Qiao L., Zhang J., Wang Z., Wu Y., Shen H., Sun M., Liu W., Xue W., Fan Y., Zhang Q., Chen B., Wu X., Shi Y., Lu F., Zhao Y., Xiao Z, & Dai J. (2022). Transplantation of neural stem progenitor cells from different sources for severe spinal cord injury repair in rat. Bioactive Materials, 23, 300-313.

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Cell Line Culture and Lentiviral Transduction

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All cell lines were cultured at 37°C with 5% CO2 in tissue culture incubators. HEK293T (female), HeLa (female), and U2OS (female) cells were cultured in Dulbecco’s modified eagle medium (DMEM) in 10% FBS (HyClone or VWR), 100 units/mL streptomycin, 100 μg/ml penicillin, and 2 mM glutamine. K562 (female) cells were maintained in RPMI-1640 with 25 mM HEPES and 2.0 g/L NaHCo3 in 10% FBS, 2 mM glutamine, 100 units/mL streptomycin, and 100 mg/mL penicillin. WTC Gen1c iPSCs (male) were cultured in mTESR media (STEMCELL Technologies) under feeder-free conditions on growth factor-reduced Matrigel (BD Biosciences). Cells were passaged using Accutase (STEMCELL Technologies) and seeded on Matrigel coated plates with mTESR media supplemented with p16-Rho-associated coiled-coil kinase (ROCK) inhibitor Y-27632 (10 μM; Selleckchem).
Lentiviral particles were produced by transfecting standard packaging vectors into HEK293T using TransIT-LT1 Transfection Reagent (Mirus, MIR2306). Media was changed 24 hours post-transfection with complete DMEM supplemented with 15 mM HEPES. Viral supernatants were harvested 48–60 hours after transfection and filtered through a 0.45 μm PVDF syringe filter. Lentiviral infections included polybrene (8 μg/ml).

Nuñez J.K., Chen J., Pommier G.C., Cogan J.Z., Replogle J.M., Adriaens C., Ramadoss G.N., Shi Q., Hung K.L., Samelson A.J., Pogson A.N., Kim J.Y., Chung A., Leonetti M.D., Chang H.Y., Kampmann M., Bernstein B.E., Hovestadt V., Gilbert L.A, & Weissman J.S. (2021). Genome-wide Programmable Transcriptional Memory by CRISPR-based Epigenome Editing. Cell, 184(9), 2503-2519.e17.

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Invasion Assay for Prostate Cancer

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The invasion assay was conducted using a 24-well trans-well chamber with a polycarbonate membrane with a pore size of 8 μm (Corning, NY, USA). The membrane was coated with 60 μL of 17% Matrigel (BD Sciences, San Jose, CA, USA) and serum-free RPMI 1640 medium to form a matrix barrier. After the Matrigel was allowed to solidify at 37 °C for 2 h, 22Rv1 and LNCaP cells (100,000 cells/mL, treated with mitomycin C (10 μg/mL, Selleck, Houston, TX, USA) 2 h for inhibiting cell proliferation) that had or had not been co-cultured with M2 macrophage were added to the upper compartment of the chamber; the lower chamber was filled with 0.6 mL of medium supplemented with 10% FBS as a chemoattractant. After incubation at 37 °C for 24 h, Matrigel and the cells remaining in the upper chamber were removed using cotton swabs. Cells on the lower surface of the membrane were fixed in 4% paraformaldehyde and stained with Giemsa (Solarbio, Beijing, China). Cells in 3 microscopic fields were counted and photographed. All experiments were performed in triplicate.

Zheng T., Ma G., Tang M., Li Z, & Xu R. (2018). IL-8 Secreted from M2 Macrophages Promoted Prostate Tumorigenesis via STAT3/MALAT1 Pathway. International Journal of Molecular Sciences, 20(1), 98.

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Routine Culture and Maintenance of hPSCs

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For routine culture and maintenance, hESC lines (H9 and H1, WiCell) and iPSC lines (TMOi001-A, ThermoFisher) were grown on Matrigel (Corning, lot #1313001) coated plates in mTeSR1 medium (STEMCELL Technologies) and 1% penicillin/streptomycin (Life Technologies) at 37°C and 5% CO₂ with daily media changes. For single-cell seeded maintenance or experiments, cells were singularized with Accutase (STEMCELL Technologies) at 37°C for 4 min and seeded onto Matrigel-coated plates in media containing 10 μM ROCK inhibitor Y-27632 (Selleckchem). 6-well plates were utilized for hPSC maintenance while hPSC differentiations were performed on 24-well plates for the 2D conditions. Cells were seeded at a density of 15k cell/cm² for maintenance or 35k cells/cm² for differentiations (see hPSC differentiation in 2D). For cluster seeded maintenance or experiments, hESCs were collected with Versene (ThermoFisher) at 37°C for 4 min and were dissociated with a P1000 wide bore pipette (ThermoFisher). Maintenance passages ranged from 1:10-1:12 (∼15k cells/cm²). Cell counts were determined by further trituration and counting on a hemocytometer.

Johnson H.J., Chakraborty S., Muckom R.J., Balsara N.P, & Schaffer D.V. (2022). A scalable and tunable thermoreversible polymer for 3D human pluripotent stem cell biomanufacturing. iScience, 25(10), 104971.

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