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Manufactured by Atlanta Biologicals

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Corning (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Voltohmmeter, by World Precision Instruments (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Amphotericin b, by Merck Group (1 mentions) Gentamicin, by Merck Group (1 mentions) G418 sulfate, by MP Biomedicals (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions)

4 protocols using premium select

Differentiation of HEK and SH-SY5Y cells

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A HEK cell line stably transfected with human TrkB was a gift from M. V. Chao (New York University). Cells were cultured in a 5% CO 2 atmosphere at 37°C in Dulbecco's modified Eagle's medium (DMEM) with GlutaMAX (Life Technologies) supplemented with 10% fetal bovine serum (FBS) (Premium Select, Atlanta Biologicals), penicillin (100 U/ml) and streptomycin (100 mg/ml) (Life Technologies), and G418 sulfate (200 mg/ml) (MP Biomedicals).
SH-SY5Y cells were cultured in DMEM and Nutrient Mixture F-12 (HAM) [DMEM/F-12 (1:1)] (Life Technologies) supplemented with 10% FBS (Premium Select, Atlanta Biologicals), penicillin (100 U/ml), and streptomycin (100 mg/ml) (Life Technologies). To perform the experiments, cells were seeded in 24-well plates (2 × 10 5 cells per well) and incubated for 48 hours, and then the cells were washed with phosphatebuffered saline (PBS) and incubated for 48 hours in DMEM/F12 (1:1) supplemented with 4% FBS, penicillin (100 U/ml), streptomycin (100 mg/ml), and 10 mM all-trans-retinoic acid (Sigma). Before treatment with compounds, cells were incubated for at least 6 hours in medium without all-trans-retinoic acid.

Boltaev U., Meyer Y., Tolibzoda F., Jacques T., Gassaway M., Xu Q., Wagner F., Zhang Y.L., Palmer M., Holson E, & Sames D. (2017). Multiplex quantitative assays indicate a need for reevaluating reported small-molecule TrkB agonists. Science signaling, 10(493).

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MDA-MB-231 Cell Stimulation Protocol

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The human breast cancer cell line, MDA-MB-231, was maintained in Dulbecco’s minimal essential medium (DMEM) (Corning cellgro), supplemented with 10% fetal calf serum (FCS, Atlanta Biologicals Premium Select Heat inactivated at 56°C, Cat#S11550H Lot # H12123) and 1 × L-glutamine, penicillin, and streptomycin (Gibco). To prepare MDA-MB-231 cells for stimulation, the cells (1 × 10⁶ cells/ml in DMEM/10% FCS) were incubated in polystyrene round tubes at 37 °C for 15 min. The cells were centrifuged at 1,200 × g for 5 min, and resuspended in DMEM/10% FCS containing 2 ng/ml IFNγ. The cells were stimulated for 10 min at 37 °C, centrifuged, washed once in PBS to stop the reaction, and prepared for FACS analysis.

Matsunaga K.I., Kimoto M., Hanson C., Sanford M., Young H.A, & Hirao I. (2015). Architecture of high-affinity unnatural-base DNA aptamers toward pharmaceutical applications. Scientific Reports, 5, 18478.

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Isolation and Characterization of Human Adipose-Derived Stem Cells

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Excess adipose tissue was collected from five female premenopausal donors (ages 24 to 36) in accordance with an approved IRB protocol at UNC Chapel Hill (IRB 04-1622) [13 (link)]. Human ASC were isolated from the tissue as previously described by our lab and others [14 (link)–16 (link)]. Cells were expanded in complete growth medium (CGM) comprised of alpha-modified minimal essential medium (α-MEM with L-glutamine) (Invitrogen, Carlsbad CA), 10% fetal bovine serum (FBS) (Premium Select, Atlanta Biologicals, Lawrenceville GA), 200 mM L-glutamine, and 100 I.U. penicillin/100 μg/mL streptomycin (Mediatech, Herndon VA). The cells were cultured at 37°C in 5% CO₂ until reaching 80% confluency and then passaged using trypsin-EDTA (Invitrogen). A superlot was generated by pooling equal numbers of cells from the five individual donor cell lines into a single culture vessel and characterized for multilineage differentiation potential, ensuring the cells differentiated representative of an average of the five cell lines [13 (link)].

Mellor L.F., Huebner P., Cai S., Mohiti-Asli M., Taylor M.A., Spang J., Shirwaiker R.A, & Loboa E.G. (2017). Fabrication and Evaluation of Electrospun, 3D-Bioplotted, and Combination of Electrospun/3D-Bioplotted Scaffolds for Tissue Engineering Applications. BioMed Research International, 2017, 6956794.

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Caco-2 Transwell Monolayer Formation

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Caco-2 cells were seeded at an initial density of 200,000 cells/cm² in the upper chamber of 6.5 mm insert Transwell (Costar; Corning Incorporated, Kennebunk, ME.) Approximately 0.25 ml of MEM (Minimum Essential Medium Eagle with Earle’s salts and L-glutamine, Corning Cellgro #10-010-CV) containing 20 % Fetal Bovine Serum (FBS; Premium Select, Atlanta Biologicals), 100 mM Sodium Pyruvate (ThermoFisher), Pen/Strep (Hyclone), Amphotericin B (Sigma) and Gentamicin (Sigma). The bottom well contained 0.5 ml MEM. The cells were incubated in a CO₂ incubator at 37°C for 5 days, with a change of medium every second day until they formed a high transepithelial resistance (TER) monolayer of 500 Ω×cm² or higher. TER was measured using a voltohmmeter (World Precision Instruments) where the measured resistance in Ohms was multiplied by the area of the Transwell filter (0.33 cm²).

Stewart T., Koval W.T., Molina S.A., Bock S.M., Lillard JW J.r., Ross R.F., Desai T.A, & Koval M. (2017). Calibrated flux measurements reveal a nanostructure-stimulated transcytotic pathway. Experimental cell research, 355(2), 153-161.

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