Effect of Shh and BM-MSC Synergism on the Proliferation of Hematopoietic Stem Cells

Abstract

To study the effect and mechanism of shh and mesenchymal stem cell（MSC）synergism on the proliferation of hematopoietic stem cells in noninvasive co-culture system in vitro. The mesenchymal stem cells were cultured in vitro，CD34+ cells were sorted by mini MACS magnetic bead separator，flow cytometry was used to identify the purity of 2 cells. CD34+ cells and MSCs were seeded to upper and low of transwell respecibely for non-contact coculture，and add exogenous shh protein for intervenece. The number of MSCs and HSCs，the total amount of RNA，the expression of ki67 and Tie-2 mRNA of HSC，the expression of VEGF and Ang-1 mRNA of MSC were detected for investigating the condition of cell proliferation and the expression of angiogenic factors. The total number of cells，the total amount of RNA and the relative expression of ki67, Tie-2, VEGF and Ang-1 in non-contact co-culture group increased and showed the following trends on the 7th day：the above-mentioned indexes in group MSC + HSC, group shh + HSC were higher than those in group HSC, while those in MSC + shh + HSC Group was higher than those in MSC + HSC and shh + HSC group. Angiogenic factors help MSC to proliferate HSC and amplify the CD34+ hematopoietic stem/progenitor cells by shh and MSC synergism in vitro coculture system which may be related with angiogenic factors.