Stem cell differentiation into angiogenesis lineage

Abstract

AIM: Fibroblast or fibroblast growth factors (FGFs), are a family of growth factors involved in angiogenesis, which is an important process in growth and development, as well as in wound healing. The aim of this study was to evaluate the possibility to differentiate human dental pulp mesenchymal stem cells into angiogenesis lineage. Their differentiation ability was evaluated with an inducer, human FGF1 transfection. MATERIALS AND METHOD: A strain of stem cell was isolated from dental pulp, expanded and further characterized. Human FGF1 was chosen to trigger it to differentiate to angiogenesis. The gene was cloned and recombinant as pcDNA3.1/V5-His-TOPO-FGF1 vector. pcDNA3. 1/V5-His-TOPO-FGF1 vector was transfected into stem cell as well as three different negative controls, blank cell without transfection, empty vector and non-related green florescent protein (GFP) vector. After 48 hours, the mRNA level of both hepatocyte growth factor (HGF) and urokinase-type plasminogen activator (uPA) were measured by real-time polymerase chain reaction for confirm angiogenesis differentiation. RESULTS: The transcription levels of HGF and uPA mRNA were significantly higher than all three negative controls after 48 hours transfection. The relative levels of HGF mRNA were 4.078 for blank cell, 12.58 for empty vector, 17.97 for non- related GFP vector and 48.59 for pcDNA3.1/V5-His-TOPO-FGF1 vector. The relative transcription levels of uPA were significantly higher than all three negative controls. The relative levels of uPA mRNA were 0.171 for the blank cell, 3.088 for the empty vector, 3.762 for the non-related GFP vector and 10.15 for the pcDNA3.1/V5-His- TOPO-FGF1 vector. CONCLUSION: The expression of markers of angiogenesis HGF and uPA is an indication of successful differentiation of human pulp stem cells to the angiogenic lineage. The protocol presented above is effective to trigger angiogenesis from dental pulp stem cells.