L glutamine l glu

In this experimental study, mouse ESCs (mESCs, C571) and PCL were gifted from the Stem Cell Technology Research Center (Iran) based on the Ethical Committee approval of the Faculty of Veterinary Medicine, Tabriz Branch, Islamic Azad University, Tabriz, Iran. Knockout Dulbecco’s modified eagle medium (DMEM-KO), Roswell Park Memorial Institute-1640 (RPMI-1640), Iscove’s modified Dulbecco’s media (IMDM) and ESC grade fetal bovine serum (FBS) were obtained from Sigma. Sterile gelatin (0.1%), LIF, L-glutamine (L-Glu), 2-mercaptoethanol (2-ME), nonessential amino acids (NEAA), penicillin and streptomycin (Pen/Strep) were purchased from Gibco (USA). SCF, IL-3, IL-6 and Fms-related tyrosine Kinase ligand (Flt3) ligand (FL) were obtained from Genescript (USA).

A mouse spleen-derived stromal cell line (mSSC) expressing yfp was generated as previously described^{4 (link)}. F9-RARE-LacZ cell line was previously described^{37 (link)}. Cells were cultured at 37 °C, 5% CO₂ in DMEM (Gibco) supplemented with 10% heat-inactivated FBS (Euroclone), 2 mM l-glutamine (l-Glu; Gibco), 100 U/ml penicillin and 100 μg/ml streptomycin (Pen/Strep; Gibco). All cell lines are monthly tested for mycoplasma. For experiments in Vitamin A deficient medium, the FBS serum was substituted with B27 supplement normal and without retinyl acetate (B27-normal and B27-vitA^-; Invitrogen). Stromal cells were treated for 24 or 48 h with different stimuli: 1 μM all-trans Retinoic Acid (in DMSO; Sigma-Aldrich), 1 μM BMS493 (in DMSO; Tocris Bioscience)^{30 (link)} or vehicle (DMSO).

The human colon colorectal adenocarcinoma (Caco-2) cell line was obtained from the American Type Culture Collection (ATCC) (USA). The cells were maintained in minimum essential medium eagle (MEM) (Sigma) supplemented with 10% heat inactivated fetal bovine serum (FBS) (Gibco Life Technologies), 100 U/ml Penicillin/Streptomycin (Gibco Life Technologies), 100 IU/ml nonessential amino acid (NEAA) (Gibco Life Technologies, Paisley, UK), and 2 mM L- glutamine (L-Glu) (Gibco Life Technologies) (termed complete cell culture medium) at 37 °C and 5% CO₂. The cells were sub-cultured using trypsin-EDTA (Gibco Life Technologies).