Differentiation MicroRNAs Affect Stemness Status of USSCs.
BACKGROUND
MicroRNAs are endogenous non-coding RNAs with important regulatory and cell fate functions. Many studies have shown that several microRNAs are obviously up-regulated during stem cell differentiation. The question rises here is weather inhibiting differentiation will affect the stemness and self renewal status of stem cells.


METHODS
miRCURY ™LNA microRNA inhibitor (anti-miR-145 and anti-let7g) are a sequence-specific and chemically modified oligonucleotide that specifically target and knockdown miR-145 and let7g miRNA molecules. Unrestricted somatic stem cells (USSCs) were isolated from umbilical cord blood and treated with LNAs. The effect of anti-miRNA transfection was assessed by quantitative real-time PCR.


RESULTS
Real-time PCR showed that LNA was efficiently introduced into the cells and reduced miR145 and Let7g expression levels to 40% and 10% in relation to corresponding scramble control, respectively. Gene expression analysis as to self renewal and expansion showed more than 3.5 fold up regulation in Oct4 in cells treated with mir145 inhibition. Similarly a significant up to 2.5 fold up-regulation in Oct4 and cMyc expression was observed in samples treated with anti-let7g.


CONCLUSIONS
Suppression in differentiation inducing microRNAs (miR-145 and let7g) can enhance the self renewal and stemness status of USSCs at transcriptional level.
Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, Tehran, Iran.
F
F
Jamshidi Adegani
L
L
Langroudi
E
E
Arefian
M
M
Soleimani