Effect of isolation methodology on stem cell properties and multilineage differentiation potential of human dental pulp stem cells
Effect of isolation methodology on stem cell properties and multilineage differentiation potential of human dental pulp stem cells
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DOI：
10.1007/s00441-013-1630-x
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发表时间：
2013-07-01
影响因子：
3.6
通讯作者：
Bronckaers, A.
中科院分区：
文献类型：
--
作者：
Hilkens, P.;Gervois, P.;Bronckaers, A.
牙髓干细胞（DPSCs）是一种有吸引力的间充质干细胞（MSC）替代来源，因为与其他更具侵入性的间充质干细胞来源的采集方法相比，它们的分离简单。然而，对于获得DPSC培养物的分离方法仍在讨论中。本研究比较了两种最广泛适用的分离方法获得的DPSCs的干细胞特性和多系分化潜力。采用髓组织酶切法（DPSC-EZ）或外植体法（DPSC-OG）分离DPSCs，在所有实验和两种分离方法中保持培养基不变。对DPSC-EZ和DPSC-OG干细胞特性的评估显示，两组之间在增殖率和集落形成方面没有显著差异。表型分析显示，DPSC-EZ和DPSC-OG均表达CD29、CD44、CD90、CD105、CD117和CD146，差异无统计学意义。通过标准免疫（组织/细胞）化学染色和透射电子显微镜下的深入超微结构分析，证实了两种干细胞类型的多系分化潜力。我们的研究结果表明，尽管两种干细胞群体的成脂分化不完全，但DPSC-EZ和DPSC-OG都能成功分化为成脂细胞、慢性成脂细胞和成骨细胞。结果表明，酶消化法和体外生长法均可获得适合骨和软骨组织替代治疗的自体DPSC资源。
Dental pulp stem cells (DPSCs) are an attractive alternative mesenchymal stem cell (MSC) source because of their isolation simplicity compared with the more invasive methods associated with harvesting other MSC sources. However, the isolation method to be favored for obtaining DPSC cultures remains under discussion. This study compares the stem cell properties and multilineage differentiation potential of DPSCs obtained by the two most widely adapted isolation procedures. DPSCs were isolated either by enzymatic digestion of the pulp tissue (DPSC-EZ) or by the explant method (DPSC-OG), while keeping the culture media constant throughout all experiments and in both isolation methods. Assessment of the stem cell properties of DPSC-EZ and DPSC-OG showed no significant differences between the two groups with regard to proliferation rate and colony formation. Phenotype analysis indicated that DPSC-EZ and DPSC-OG were positive for CD29, CD44, CD90, CD105, CD117 and CD146 expression without any significant differences. The multilineage differentiation potential of both stem cell types was confirmed by using standard immuno(histo/cyto)chemical staining together with an in-depth ultrastructural analysis by means of transmission electron microscopy. Our results indicate that both DPSC-EZ and DPSC-OG could be successfully differentiated into adipogenic, chrondrogenic and osteogenic cell types, although the adipogenic differentiation of both stem cell populations was incomplete. The data suggest that both the enzymatic digestion and outgrowth method can be applied to obtain a suitable autologous DPSC resource for tissue replacement therapies of both bone and cartilage.