Medium 199

Mammalian cell culture media PAOSMC

Experiment
Mammalian cell culture media PAOSMC
Product
Medium 199 from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
Supplement medium with 15% fetal bovine serum with penicillin [100 U/mL], streptomycin [100 U/mL], and amphotericin [0.27 μg/mL]

Publication protocol

Vascular smooth muscle cells were isolated from medial layer explants of thoracic aorta from 3-month old female pigs and were cultured in standard growth medium ([GM] M-199/15% fetal bovine serum with penicillin [100 U/mL], streptomycin [100 U/mL], and amphotericin [0.27 μg/mL]). Medium was replaced every 2–3 days. The cellular phenotype of all cells examined was confirmed by positive immunofluorescence staining with antibodies against α-smooth muscle actin as well as smooth muscle myosin heavy chain-2. Stocks of cells were stored in liquid nitrogen, and cells between passages three and six were used for study. Each experiment was initiated by seeding cells in GM into 24-well culture plates and in 4-chamber slides at 1.3×104 cells/cm2. Cells were cultured until nearly confluent (day 0), then divided into three experimental groups differing only in treatment: 1) media alone, no treatment (control); 2) CNPs, ten nephelometric turbidity units per cm2; or 3) 2 mM P as a positive control for calcification.Cells were cultured for up to an additional 28 days before harvesting.

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Papers

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Paper title
Calcifying nanoparticles promote mineralization in vascular smooth muscle cells: implications for atherosclerosis
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Medium 199 below.

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