Tryple express enzyme no phenol red

Chlorpromazine, Kolliphor EL, Miglyol 840, dimethyl sulfoxide, glycerol, propylene glycol, polyvinyl alcohol, and bovine serum albumin were purchased from Sigma-Aldrich (St. Louis, MI, USA). Lauroglycol FCC, Transcutol HP, Labrafil 1944, and Labrasol were gifted by Gattefossé (Lyon, France). The human nasal epithelial cell line (RPMI 2650) was from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). The MTT reagent 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and the buffer solutions, such as Hank’s Balanced Salt Solution (HBSS) and phosphate-buffered saline (PBS), were purchased from Sigma-Aldrich (St. Louis, MI, USA). The RPMI cell culture maintenance medium solution, TrypLE™ Express Enzyme (no phenol red), was ordered from Thermo Fisher Scientific (Waltham, MA, USA). Ninety-six-well cell plates and culturing flasks were obtained from VWR International (Debrecen, Hungary).

Male mice of different ages were euthanized using carbon dioxide asphyxiation, then their urogenital tract and right kidneys were removed. Under a microscope, microdissection was performed to isolate the prostate and bladder from the urogenital tract. Tissues were mechanically dissociated with a razor blade then enzymatically dissociated in RPMI 1640 (Gibco) containing 10% fetal bovine serum (FBS), 1 mg/mL collagenase type I (Gibco), and 0.1 mg/mL DNase (Sigma) at 37°C on a nutating platform for 60–90 minutes. After washing the cell pellets with 1X DPBS (Gibco), cells were resuspended in 37°C TrypLE Express Enzyme, no phenol red (Thermo Fisher Scientific) for 5 minutes before being quenched by adding RPMI 1640. Cells were further disrupted using the shear stress generated by drawing through an 18G syringe. Finally, cells were sequentially passed through 100 μm and 70 μm cell strainers (Corning) to produce a single-cell suspension.

Human recombinant insulin, pepsin (≥400 unit/mg protein), and pancreatin (≥3× USP specifications) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Sodium alginate was purchased from BÜCHI Labortechnik AG (Flawil, Switzerland). Calcium chloride dihydrate was ordered from VWR International (Debrecen, Hungary). Labrasol ALF (Caprylocaproyl Prolyoxyl-8-glycerides) and Labrafil M2125 CS (Linoleoyl Polyoxyl-6 glycerides) were purchased from Gattefossé (Saint-Priest, France). The Caco-2 cell line was obtained from the European Collection of Cell Cultures (ECACC, Public Health England, Salisbury, UK). MTT dye (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), phosphate buffered saline (PBS) buffer solution, Dulbecco’s Modified Eagle’s Medium (DMEM), heat-inactivated fetal bovine serum (FBS), L-glutamine, non-essential amino acids solution, and penicillin-streptomycin solution were obtained from Sigma-Aldrich (St. Louis, MO, USA). TrypLE™ Express Enzyme (no phenol red) and Pierce™ Detergent Compatible Bradford Assay Kit were ordered from Thermo Fisher Scientific (Waltham, MA, USA).