Tomlinson Matthew J, Dennis Caitriona, Yang Xuebin B, Kirkham Jennifer
Department of Oral Biology, School of Dentistry, University of Leeds, Leeds, LS9 7TF, UK,
Cell Tissue Res. 2015 Aug;361(2):529-40. doi: 10.1007/s00441-014-2106-3. Epub 2015 Feb 1.
The cell surface hydrolase tissue non-specific alkaline phosphatase (TNAP) (also known as MSCA-1) is used to identify a sub-population of bone marrow stromal cells (BMSCs) with high mineralising potential and is found on subsets of cells within the dental pulp. We aim to determine whether TNAP is co-expressed by human dental pulp stromal cells (hDPSCs) alongside a range of BMSC markers, whether this is an active form of the enzyme and the effects of culture duration and cell density on its expression. Cells from primary dental pulp and culture expanded hDPSCs expressed TNAP. Subsequent analyses revealed persistent TNAP expression and co-expression with BMSC markers such as CD73 and CD90. Flow cytometry and biochemical assays showed that increased culture durations and cell densities enhanced TNAP expression by hDPSCs. Arresting the hDPSC cell cycle also increased TNAP expression. These data confirm that TNAP is co-expressed by hDPSCs together with other BMSC markers and show that cell density affects TNAP expression levels. We conclude that TNAP is a potentially useful marker for hDPSC selection especially for uses in mineralised tissue regenerative therapies.
细胞表面水解酶组织非特异性碱性磷酸酶（TNAP）（也称为MSCA-1）用于鉴定具有高矿化潜力的骨髓基质细胞（BMSC）亚群，并且在牙髓内的细胞亚群中发现。我们旨在确定TNAP是否与人牙髓基质细胞（hDPSC）共同表达一系列BMSC标志物，这是否是该酶的活性形式，以及培养持续时间和细胞密度对其表达的影响。来自原代牙髓和培养扩增的hDPSC的细胞表达TNAP。随后的分析显示TNAP持续表达并与BMSC标志物如CD73和CD90共同表达。流式细胞术和生化分析表明，延长培养时间和增加细胞密度可增强hDPSC的TNAP表达。阻断hDPSC细胞周期也增加了TNAP表达。这些数据证实TNAP与其他BMSC标志物共同由hDPSC表达，并表明细胞密度影响TNAP表达水平。我们得出结论，TNAP是hDPSC选择的潜在有用标志物，特别是在矿化组织再生疗法中的应用。
