Detection method of gene expression of molecular markers characterizing osteogenic fifferentiation of stem cells specific for animal model of a horse, panel of primers for making the method and use of such a primer panel

摘要

Panel of primers and method of detection of gene expression of molecular markers characterising osteogenic differentiation specific to the equine animal module,where an isolated DNA from specifically differentiated equine mesenchymal cells to osteocytes (bone cells) is produced by a polymer a se chain reaction (PCR) using a primer panel comprising:at least one of the oligonucleotide prime pairs: OMD F: 5 -CAC AAC AAA CTG AAG CAA GCA -3 (21 -mer), OMD R: 5 -TGA GGG AAG CAG AGG AAA ATG -3 (21 -mer),for the detection of the gene sequence osteomodulin (OMD); ON F: 5 -GAGAGAAGGTGCAGCAATGA -3 (20 -mer), ON R: 5 -GCT TCT CGG TCA GAA GGT TG -3 (20 -mer),for the detection of the gene sequence osteonectin (ON); RUNX2 F: 5 -CCC CAA GTA GCC ACC TAT CA -3 (20 -mer), RUNX2 R: 5 -GCCTGGTCTGTSSTCTGA -3 (20 -mer),for the detection of the gene sequence of the transcription factor RUNX2 (RUNX2); OPN F: 5 -CTG ATT CCA ACG TT -3 (20 -mer), OPN R: 5 -ATG TCG TTG CTT TCC ACA GG -3 (20 -mer),for the detection of the gene sequence of osteopontin (OPN) and subsequently electrophoresis of PCR reaction products shall be performed.The solution allows rapid, simple and specific detection of cells differentiated into osteocytes using specific markers in cell isolates especially in the animal species of the horse.