GSM6310583: BVE, control 2; Mus musculus; RNA-Seq
Thyroid tumors were collected aseptically from donor mice (BVE and BVE-Ctnnb1null) using blunt dissection, mechanically dissociated by mincing and passing through a 40-μM mesh sterile screen, and suspended in DMEM/F12 growth medium (10% fetal bovine serum, 100 units/ml penicillin, 100 ug/ml streptomycin). Cells were cultured in DMEM/F12 growth medium containing 4 mU/mL bovine TSH (Sigma-Aldrich). For establishment of immortalized cell lines, cells were cultured continually for more than 6 months and passaged weekly or bi-weekly at 1:3 ratio with 0.25% trypsin. BVE-Ctnnb1wt (BVE) and BVE-Ctnnb1null cell lines had about 25 and 20 passages, respectively before they were used for experiments.  Thyroid origin of the cell lines was confirmed by genotyping to rule out fibroblast contamination. Genotyping of the cell lines and expression of genes for thyroid hormone synthesis were verified every 6 months when the cells were in culture. Libraries were constructed using an Illumina (San Diego, Ca, USA) TruSeq RNA Library Prep kit according to the manufacturer's procedure.