Distearoyl sn glycerol 3 phosphocholine dspc

Cholesterol was purchased from Sigma Aldrich, while distearoyl-sn-glycerol-3-phosphocholine (DSPC) and PEG2000-DMG were obtained from Avanti Polar Lipids. Caco-2 cells were purchased from the American Type Culture Collection. Minimum Essential Media (MEM), trypsin, penicillin/streptomyocin, phosphate buffered saline (PBS), and fetal bovine serum (FBS) were purchased from Life Technologies. Antibodies reactive to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and alpha tubulin were purchased from Abcam. Silencer Select siRNA for GAPDH and green flourescence protein (GFP) were orderd from Life Technologies.

Nanoparticles were formulated with the three‐tailed version of lipidoid 306O₁₃,36 which was synthesized via the Michael addition of 3,3′‐diaminodipropylamine (Acros Organics) to tridecyl acrylate (Pfaltz and Bauer) at a stoichiometric ratio of 1:3 as described previously.37, 39 The lipidoid was then purified over a silica column on a Teledyne ISCO chromatography system to isolate the three‐tailed product. For nanoparticle formulation, a lipid solution was formed by mixing the lipidoid, distearoyl‐sn‐glycerol‐3‐phosphocholine (DSPC, Avanti Polar Lipids, Alabaster, Alabama), cholesterol (Sigma Aldrich), and C14‐PEG (Avanti Polar Lipids) at a molar ratio of 50:10:38.5:1.5 in ethanol (Sigma Aldrich) and 10 mM citrate buffer. Silencer Select Pre‐Designed siRNA against TNFα (s75248, Thermo Fisher) was diluted in 10 mM sodium citrate and combined with the lipid solution at a final lipidoid to siRNA weight ratio of 5:1. The solution was vortexed after each reagent addition, and the lipid solution was added to the siRNA solution before being diluted to the desired final concentration in phosphate buffered saline (PBS).

Cholesterol was purchased from Sigma Aldrich, while distearoyl-sn-glycerol-3-phosphocholine (DSPC) and PEG2000-DMG were obtained from Avanti Polar Lipids. Dual HeLa and Caco-2 cells were purchased from American Type Culture Collection (Manassas, VA). Porcine pepsin, pancreatin, bile salts, and mucin type II were obtained from Sigma (St Louis, MO). Dulbecco’s Modified Eagles Media (DMEM), trypsin, penicillin/streptomyocin, phosphate buffered saline (PBS), fetal bovine serum (FBS), and GAPDH siRNA were purchased from Thermo Fisher (Waltham, MA). Corning BioCoat HTS 1 µm porous support Transwell plates, basal seeding medium (BSM), Corning EnteroSTIM enterocyte differentiation medium (EDM), and MITO + serum extender were obtained from VWR (Radnor, PA). Anti-firefly luciferase siRNA (siLuc) was purchased from Dharmacon (Lafayette, CO).