Transcription profiling of mouse thymus glandwfrom normal and ollagen induced arthritis animals

ABSTRACT: DBA/1J and DBA-2/J mice (12-14 weeks old) weighing 18�22 g were housed in temperature-controlled rooms (22�25�C) in the animal facility of the School of Medicine of Ribeir�o Preto, University of S�o Paulo, S�o Paulo, Brazil, and received water and food ad libitum. DBA/1J and DBA-2/J male mice were used in type II collagen immunizations. For CIA, at day zero, DBA/1J males were intradermally injected at the base of the tail with 200 �g of bovine type II collagen (CII) (Sigma) in 100�l of 0.05 M acetic acid emulsified with equal volume of complete Freund�s adjuvant (Chondrex, Redmond, WA). On day 21, a second injection of CII (200 �g in acetic acid) was given intraperitoneally (i.p). DBA/2J males were also immunized with CII and served as controls. A caliper was used to determine the paw diameter and swelling was determined as the increase in diameter compared to day zero of immunization. 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 and from 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).

ORGANISM(S): Mus musculus

DISEASE(S): collagen induced arthritis

SUBMITTER: Geraldo Passos 

PROVIDER: E-MEXP-2338 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress