MicroRNA profiling of human umbilical cord mesenchymal stem cells during osteogenic differentiation
ABSTRACT: The time course experiment was designed as follow: (1) Mesenchymal stem cell (indifferentiated cells)/ time 0h; (2) After 24 h of osteogenic induction; (3) After 48 h of osteogenic induction (4) After 7 days of osteogenic induction.
Project description:The current project is within the range of molecular immunogenetic auto immune diseases and refers to the comparative study of promiscuous gene expression of tissue-specific antigens (TSAs) in the thymus of NOD mice line (non obese diabetic) who plays the auto-immune diabetes mellitus type 1, during the transition from state pre-diabetics to diabetics. Use the technology of oligo arrays to investigate the expression of miRNAs and cDNA microarrays to investigate the expression of genes encoding the messenger RNAs including TSAs (tissue specific antigens).
Project description:Umbilical cord mesenchymal stem cells were stimulated to osteogenesis by 24h, 48h and 7 days. Undifferentiated and treated cells have their total RNA extracted and 25 ug were used to extract micro RNAs by Flash PAGE Fractionator (Ambion). These miRNA were labeled and used to hybridize to 662 oligo arrays corresponding to miRNA sequences from human, rat and mouse. The hybridized slides were scanned in the Amersham Automatic Slide Processor, quantified by TIGR Spotfinder software and submitted to stathiscal treatment in TIGR MEV software.
Project description:Small interfering RNA ( siRNA) was used to knockdown Autoimmune regulator (Aire) gene by in vivo electroporation of the thymus of BALB-c mice. In this set of data we include control and Aire-knockdown mTEC cells isolated from thymus of BALB-c mice.
Project description:The aim of this study is to characterize how the extracellular matrix secreted by adipose-derived stem cells (ADSC) during osteogenesis affects the differentiation process. Specifically, ADSC undergo osteogenesis by following similar maturational phases as bone marrow-derived stem cells. However, it is unclear how the differentiation process is the same and how it differs. We first focused on ADSC behavior by analyzing whole transcriptome changes in response to osteogenic media supplements added into the tissue culture medium. We then developed osteogenic differentiation expression profiles for the ADSC and identify key genes and pathways that serve as an osteogenesis signature. This expression signature acted as a template for comparison of how extracellular matrix (ECM) affected ADSC differentiation. We studied ADSC induced to differentiate on ECM isolated from day 16 in the differentiation process (the midpoint in osteogenesis) as well as ECM from day 11 in the differentiation process. Ultimately, we aim to dissect the relationship between cells grown on ECM as an in vitro growth substrate and cells grown on tissue culture plastic; both in the presence of osteogenic supplements. This study, will allow us to determine the extent to which ECM affects the differentiation process.
Project description:Hypoxia results in the changes in expression of many genes, the majority of which are mediated via the transcriptional activity of the hypoxia inducible factor (HIF) complex. However, other mechanisms of gene regulation by hypoxia are likely and include control of mRNA stability, regulation of mRNA translation and regulation mediated by micrornas. The aim of this study is to identify microRNAs which expression is regulated by hypoxia. We chose the breast cancer line MCF7 for study as we had previously characterised the expression of the components of the HIF system in that cell line and undertaken an extensive study of the gene expression profile in response to hypoxia, a prolyl hydroxylase inhibitor dimethyloxalylglycine and HIF-1a isoform manipulations (Eldvidge. G.P. et al. (2006) JBC, vol. 281, 22, 15215-15226).