Human bone marrow stromal (skeletal) stem cells (hBMSC) are cells that retain a multi-lineage differentiation potential and are thus increasingly being investigated for use in clinical applications. In vivo BMSC, which comprise approximately 0.1% of the bone marrow compartment, are thought to maintain their ‘stemness’ in the niche for prolonged periods of time by inducing a reversible exit of the cell cycle termed quiescence (G0). However, little is known about the induction, regulation or maintenance of this state in hBMSC. Quiescence, which usually occurs during the G1 stage of the cell cycle, allows the conservation of cells against future regenerative needs, and may, in addition, maintain equilibrium between self-renewal and differentiation. We have established and characterised induction of quiescence in an immortalised hBMSC line using cellular suspension. Proliferation markers, such as Ki67 and BRDU, were demonstrated to down regulate within 24hrs of cell suspension and reestablishment of proliferation was observed by 24hrs post G0 as demonstrated by Ki67 and BRDU staining. In addition, no staining was observed for the senescence marker b-Galactosidase during or post G0. Gene expression profiling of the cells demonstrated down-regulation of cyclin (CCNA2, CCND1, CCNE1, CCNB1) and proliferation markers (Ki67) markers during G0 and up-regulation of the osteogenic genes RUNX2 and OPN. RT-PCR analysis of osteogenic differentiation of cells post G0 demonstrated an increase in osteogenic gene expression and earlier matrix deposition in cells that had undergone G0. No difference in differentiation capacity was observed during adipogenic differentiation. In conclusion, G0 has ben established in an immortalised hBMSC line and data has demonstrated that post G0 cells have an increased capacity for OB induction.
|Status||Udgivet - 2014|
|Begivenhed||12th Annual Meeting of the International Society for Stem Cell Research - Vancouver, Canada|
Varighed: 18. jun. 2014 → 21. jun. 2014
Konferencens nummer: 12
|Konference||12th Annual Meeting of the International Society for Stem Cell Research|
|Periode||18/06/2014 → 21/06/2014|
Harkness, L., Rumman, M., Kassem, M., & Dahwan, J. (2014). Establishing quiescence in human bone marrow stem cells leads to enhanced osteoblast marker expression. Poster session præsenteret på 12th Annual Meeting of the International Society for Stem Cell Research, Vancouver, Canada.