title: GSM8680241: BVEp53; Mus musculus; RNA-Seq
library_construction_protocol: Thyroid tumors were collected aseptically from transgenic mice (harbouring BrafV600E (BVE), BVE and Trp53, KrasG12D, and KrasG12D and Cdkn2a, respectivelu) 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. To establish pulmonary metastatic thyroid cancer cell lines, 1 x 106 cells were injected to tail vain of 5 nude mice for each group. Six weeks after injection, pulmonary metastatic tumors were collected aseptically from the mice using blunt dissection, then mechanically dissociated by mincing and passing through a 40-μM mesh sterile screen, and suspended in DMEM/F12 growth medium for 3 month with a total of 6 passages to eliminate contaminated stromal fibroblasts, lymphocytes, and microphages present in the tumor cell culture. The primary cells were considered as permanent cell lines after six passages. The established metastatic cell lines were named as BVE-Met1, BVEp53-Met1 Kras-Met1, and Krasp16-Met1, respectively. They were re-injected (1 x 106 cells) to a new group of nude mice (n=5 for each group) via tail vain for enrichment of cells with high metastatic potential. Three weeks following injection, lung metastatic tumors were harvested and propagated in DMEM/Ham's F12 growth medium for 3 month with at least 6 passages. The metastatic cell lines were named as BVE-Met2, BVEp53-Met2, Kras-Met2, and Krasp16-Met2, respectively. The thyroid origin of these cell lines were confirmed by genotyping. Libraries were constructed using an Illumina (San Diego, Ca, USA) TruSeq RNA Library Prep kit according to the manufacturer's procedure. RNA-Seq on BGISEQ-500 platform