All cell lines were cultured as mono-layers and maintained in filter-capped tissue culture flasks coated with 1% (v/v) Matrigel (Matrigel GFR Basement Membrane Matrix LDEV-Free; BD Biosciences, Mississauga, Canada) in Dulbecco’s Modified Eagle Medium (DMEM; ThermoFisher, MA USA). Culture vessels were coated with Matrigel by covering the internal surface with 1% Matrigel in DMEM High Glucose Pyruvate media and incubation for 1 h at 37 °C or overnight at 4 °C. Cells were maintained at 37 °C and 5% CO2 in KnockOut™ DMEM/F-12 (ThermoFisher) supplemented with recombinant Human EGF (20 ng/mL; ThermoFisher), recombinant Human FGFb (10 ng/mL; ThermoFisher), glutamine (20 mM/mL; ThermoFisher), penicillin/streptomyocin (100 U/mL; ThermoFisher), StemPro Neural Supplement (20 ng/mL; ThermoFisher) and heparin (20 ng/mL; Sigma-Aldrich, MO USA). Cells were sub-cultured for a maximum of 35 passages, with cells detached for passaging with StemPro Accutase (ThermoFisher).
Matrigel gfr basement membrane matrix ldev free
Manufactured by BD
Sourced in Canada
Matrigel GFR Basement Membrane Matrix LDEV-Free is a solubilized basement membrane preparation extracted from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is a complex mixture of extracellular matrix proteins, including laminin, collagen IV, heparan sulfate proteoglycans, and entactin.
Automatically generated - may contain errors
Lab products found in correlation
Heparin, by Merck Group (2 mentions)
Glutamine, by Thermo Fisher Scientific (2 mentions)
Knockout dmem f 12, by Thermo Fisher Scientific (2 mentions)
Stempro neural supplement, by Thermo Fisher Scientific (2 mentions)
Recombinant human egf, by Thermo Fisher Scientific (2 mentions)
Recombinant human fgfb, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomyocin, by Thermo Fisher Scientific (2 mentions)
Stempro accutase, by Thermo Fisher Scientific (1 mentions)
2 protocols using matrigel gfr basement membrane matrix ldev free
1
Culturing Stem Cells in Matrigel-Coated Flasks
2
Culture and Maintenance of Cell Lines on Matrigel-Coated Polyacrylamide Hydrogels
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All cell lines were cultured and maintained in lter-capped tissue culture asks coated with 1 % (v/v) Matrigel (Matrigel GFR Basement Membrane Matrix LDEV-Free; BD Biosciences, Mississauga, Canada) in Dulbecco's Modi ed Eagle Medium (DMEM; ThermoFisher, MA USA). Culture vessels were coated with Matrigel by covering the internal surface with 1 % Matrigel in DMEM High Glucose Pyruvate media and incubation for 1 hour at 37°C or overnight at 4°C. Cells were maintained at 37°C and 5 % CO 2 in KnockOut™ DMEM/F-12 (ThermoFisher) supplemented with recombinant Human EGF (20 ng/mL; ThermoFisher), recombinant Human FGFb (10 ng/mL; ThermoFisher), glutamine (20 mM/mL; ThermoFisher), penicillin/streptomyocin (100 U/mL; ThermoFisher), StemPro Neural Supplement (20 ng/mL; ThermoFisher) and heparin (20 ng/mL; Sigma-Aldrich, MO USA).
Polyacrylamide (PA) hydrogels PA 35 mm Petrisoft™ (plastic-bottomed) and Softview™ (glass-bottomed) products (Matrigen, CA USA) of de ned stiffness (0.5, 8 and 50 kPa) were used for bright eld microscopy and confocal microscopy, respectively. All PAGs were coated with 1 % Matrigel solution for 1 hour and rinsed with phosphate buffered saline (PBS, ThermoFisher) before cells were seeded.
Polyacrylamide (PA) hydrogels PA 35 mm Petrisoft™ (plastic-bottomed) and Softview™ (glass-bottomed) products (Matrigen, CA USA) of de ned stiffness (0.5, 8 and 50 kPa) were used for bright eld microscopy and confocal microscopy, respectively. All PAGs were coated with 1 % Matrigel solution for 1 hour and rinsed with phosphate buffered saline (PBS, ThermoFisher) before cells were seeded.
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