Proud of Joshua DeJong and Pat as they present their research this morning at the NCUR conference at the University of Kentucky! EFFECT OF OXYGEN LEVELS AND SERUM CONCENTRATION ON MRNA EXPRESSION OF PARACRINE FACTORS IN MESENCHYMAL STEM CELLS Authors- Patrick Page, Joshua DeJong, Robert A. Boomsma Faculty mentor- Robert A. Boomsma Department- Biology Institution- Trinity Christian College Institutional address- 6601 W. College Drive, Palos Heights Illinois 60463 Previous studies have shown that mouse bone marrow mesenchymal stem cells (MSC) secrete paracrine factors that may be responsible for their beneficial effect on mouse hearts after coronary artery occlusion (Boomsma and Geenen, PLoSONE 7:e3685, 2012). The purpose of this study was to determine the effect of oxygen levels and serum concentrations on the gene expression for monocyte chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGF), and macrophage inflammatory protein-1 alpha and beta (MIP-1a, MIP-1b) by MSC during cell culture. MSC were cultured in Mesencult (Stem Cell Technologies) under normoxic (21% O2) and hypoxic (1% O2) conditions in the presence of either 5% or 20% serum supplement. RNA was isolated from the cells using RNAqueous 4 PCR (Ambion), treated with DNAse and quantified by RT-qPCR using an iScript cDNA Synthesis Kit and iQ SYBR Green Mastermix (BioRad). Relative expression was determined using the reference gene YWHAZ; no-RT and water controls were included. Mean and standard error was calculated for each group and significance was determined using student’s T-test (p
Posted on: Thu, 03 Apr 2014 12:49:44 +0000
Recently Viewed Topics
© 2015