hCMEC/D3 cells (Cat #SCC066) and media (Cat #SCME004), dish coating material type I rat collagen (Cat #08–115) were purchased from MilliporeSigma. Primary human astrocytes isolated from cerebral cortex (Cat # 1800), astrocyte medium (Cat # 1801), trypsin neutralization solution (Cat # 0113) and dish coating material poly-L-lysine (Cat # 0403) were purchased from ScienCell Research Laboratories. All cells were maintained according to manufacturer instructions. Generation and culture of hiPSC-derived neurons and astroglia from human cortical spheroids (hCS) were described previously.17 (link),26 (link),36 (link) Briefly, hiPSC derived from fibroblasts and maintained on MEFs were used to generate hCS for more than 150 days, after which they were enzymatically dissociated and immunopanned into purified neurons (Mouse anti-Thy1 [CD90], BD Biosciences, Cat. 550402) and astroglia cells (Mouse anti-HepaCAM, R&D, Cat # MAB4108) as previously described.26 (link) The isolated cells were seeded on poly-d -lysine coated plates and maintained in a Neurobasal/DMEM based serum-free medium.26 (link) All cell cultures were maintained in a humidified incubator at 37°C with 5% CO2. All human stem cell work was performed with approval from the Stanford Human Stem Cell Research Oversight committee.
Trypsin neutralization solution
Manufactured by ScienCell
Sourced in United States
Trypsin neutralization solution is a reagent used to inactivate the enzymatic activity of trypsin. Trypsin is commonly used in cell culture applications to detach adherent cells from a growth surface. The neutralization solution contains components that inhibit the proteolytic activity of trypsin, allowing the cells to be collected and subcultured.
Automatically generated - may contain errors
Lab products found in correlation
Poly d lysine, by R&D Systems (1 mentions)
Hcmec d3 cells, by Merck Group (1 mentions)
Astrocyte medium, by ScienCell (1 mentions)
Phosphate buffered saline (pbs), by ScienCell (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Poly l lysine (pll), by ScienCell (1 mentions)
Bardoxolone, by MedChemExpress (1 mentions)
Endothelial cell growth factor, by ScienCell (1 mentions)
Nrf2 small interfering rna sirna, by GenePharma (1 mentions)
Lamin b1, by Proteintech (1 mentions)
Anti mouse, by Bioworld Technology (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Anti rabbit, by Bioworld Technology (1 mentions)
Gapdh, by Proteintech (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Penicillin streptomycin, by ScienCell (1 mentions)
Fetal bovine serum (fbs), by ScienCell (1 mentions)
3 protocols using trypsin neutralization solution
1
Generation and Culture of hiPSC-Derived Neurons and Astroglia
2
Osteogenic Differentiation of hBMSCs
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Primary hBMSCs (cat. no. 7500), mesenchymal stem cell basal culture medium, PBS, poly-L-lysine, trypsin neutralization solution, trypsin/EDTA digestion buffer, mesenchymal stem cell osteogenic differentiation medium (MODM) and osteogenic staining solution (2% Alizarin red S staining solution) were purchased from ScienCell Research Laboratories, Inc. (San Diego, CA, USA). The alkaline phosphatase (ALP) staining reagent kit was purchased from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany). TRIzol® was obtained from Life Technologies (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The quantitative polymerase chain reaction (qPCR) reagent kit was supplied by Applied Biosystems (Thermo Fisher Scientific, Inc.).
3
Nrf2 Pathway Modulation in Endothelial Cells
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3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), endothelial cell medium (ECM), fetal bovine serum (FBS), penicillin/streptomycin, endothelial cell growth factor, and trypsin neutralization solution were purchased from ScienCell (San Diego, California, USA). 0.25% trypsin-EDTA was purchased from Gibco (Grand Island, NY, USA). 2′,7′-Dichlorofluorescein diacetate DCF-DA was purchased from Beyotime. Primary antibodies Nrf2, HO-1, NQO1, GAPDH, and Lamin B1 were from Proteintech (Wuhan, China). Secondary antibodies including anti-mouse and anti-rabbit were purchased from Bioworld Technology (Minnesota, USA). Bardoxolone and ML385 were purchased from MedChemExpress (Shanghai, China). The specific Nrf2 small-interfering RNA (siRNA) and negative control of siRNA were purchased from GenePharma (Shanghai, China). Enhanced chemiluminescence (ECL) western blotting detection solution was obtained from Beyotime (Shanghai, China).
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