The protocol was published [19 (link)] with minor modifications. After the cells reached confluency, approximately 80%, the cells were transferred to a plate pre-coated with 2% Matrigel (Corning, Corning, NY, Cat#: 356234) and 10 μg/mL Poly-D-lysine (Advanced BioMatrix, Carlsbad, CA, Cat#5049), in maturation complete media, which comrpised Neurobasal™ Plus Medium (Gibco™, Waltham, MA, # A3582901), with 1× B-27™ Plus (Gibco™, Waltham, MA, #A3582801), 1× Glutamax, (Gibco™, #35050061), supplemented with 50 ng/mL of β-NGF (Peprotech, Rocky Hill, NJ, Cat#450-01), 25 ng/mL of GDNF (Gibco™, #PHC7041), CNTF, and NT-3 (Peprotch, #450-13, 450-03, respectively), 1 μg/mL of Tetracycline (Sigma-Aldrich, St. Louis, MO, Cat#: T-7660) and 25 μM of Forskolin (Cayman Chemical, Ann Arbor, MI, Cat#66575-29-9), in a density of 15,544 cells/cm2 for 7 days media change for every 3–4 days.
Poly d lysine
Manufactured by Advanced BioMatrix
Poly-D-lysine is a synthetic polypeptide composed of the amino acid D-lysine. It serves as a surface coating agent to promote cell adhesion and growth in cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Tetracycline, by Merck Group (1 mentions)
Forskolin, by Cayman Chemical (1 mentions)
B 27 plus, by Thermo Fisher Scientific (1 mentions)
Neurobasal plus medium, by Thermo Fisher Scientific (1 mentions)
β ngf, by Thermo Fisher Scientific (1 mentions)
Matrigel, by Corning (1 mentions)
Acridine orange, by Thermo Fisher Scientific (1 mentions)
Facs buffer, by Thermo Fisher Scientific (1 mentions)
2 protocols using poly d lysine
1
Neuronal Maturation Protocol for Regenerative Research
2
Laminin-based Cell Culture Protocol
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Laminin (isoform 1, 90% purity, #L2020), anti-sulfatide (clone A2B5, #mab312), sodium metabisulfite (NBS, Na2S2O5,161519) were obtained from Sigma-Aldrich (St. Louis, MO). IgM negative control (#MCA692) was obtained from BioRad (Hercules, CA). Poly-d-lysine (#5049) was obtained from Advanced Biomatrix (San Diego, CA). Acridine orange (#A3568) and FACS buffer (#FC001) were obtained from Fisher Scientific (Hampton, NH). Unless otherwise indicated, reagents were reconstituted and diluted in phosphate-buffered saline (PBS).
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