| Experiment Id | E-MEXP-2339 | Name | Transcription profiling of mouse thymus gland from pre-diabetic and diabetic animals |
| Experiment Type | transcription profiling by array | Study Type | Baseline |
| Source | ArrayExpress | Curation Date | 2018-08-16 |
| description | Female NOD mice were born in specific pathogen free (SPF) conditions at the CEMIB-UNICAMP animal facility (University of Campinas, Sao Paulo, Brazil) and maintained in SPF mini-isolators in our laboratory at the University of Sao Paulo, Campus of Ribeirao Preto, Brazil. We studied both pre-diabetic (8.2 week-old) and diabetic (20.2 week-old) animals. Diabetes was confirmed by blood glucose levels (.250 mg glucose/dL) using the Accu Check Active Kit (Roche Diagnaœstica Brasil, Sao Paulo, Brazil). The thymic stroma was separated from the whole thymus. The mTEC 3.10 medullary thymic epithelial cell line was established from C57BL/6 mice, and the original medullary phenotype was confirmed by immunostaining with anti-cytokeratin monoclonal antibodies. In addition, the CD80+ phenotype was confirmed using fluorescent activated cell sorting (FACS) analysis. Cells were cultured in 10% fetal bovine serum-supplemented RPMI 1640 medium at 37oC and 5% CO2. Total RNA was extracted from 1x107 stromal cells (from pre-diabetic and diabetic animals) and 1x107 mTEC 3.10 cells using Trizol Reagent and following the manufacturer's instructions (Invitrogen, Carlsbad, CA, USA). RNA preparations were confirmed to be free of proteins and phenol using UV spectrophotometry and the state of degradation was assessed using agarose gel electrophoresis (ethidium bromide staining). |
