Department of Functional Morphology, Laboratory of Histology, Biomedical Research Institute (BIOMED), Hasselt University, Diepenbeek, Belgium.
Cell Tissue Res. 2013 Jul;353(1):65-78. doi: 10.1007/s00441-013-1630-x. Epub 2013 May 29.
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.
牙髓干细胞（DPSCs）是一种有吸引力的间充质干细胞（MSC）来源，因为与收获其他 MSC 来源相关的更具侵入性的方法相比，其分离方法更为简单。然而，用于获得 DPSC 培养物的首选分离方法仍在讨论中。本研究比较了通过两种最广泛适应的分离程序获得的 DPSCs 的干细胞特性和多能性分化潜能。DPSCs 要么通过牙髓组织的酶消化（DPSC-EZ），要么通过外植体方法（DPSC-OG）分离，同时在所有实验中和两种分离方法中保持培养基不变。对 DPSC-EZ 和 DPSC-OG 的干细胞特性的评估表明，两组之间在增殖率和集落形成方面没有显着差异。表型分析表明，DPSC-EZ 和 DPSC-OG 均对 CD29、CD44、CD90、CD105、CD117 和 CD146 的表达呈阳性，没有显着差异。两种干细胞类型的多能性分化潜能均通过使用标准免疫（组织/细胞）化学染色并通过透射电子显微镜进行深入的超微结构分析来证实。我们的结果表明，DPSC-EZ 和 DPSC-OG 都可以成功分化为脂肪细胞、软骨细胞和成骨细胞类型，尽管两种干细胞群体的脂肪细胞分化均不完全。这些数据表明，酶消化和生长方法都可以用于获得适合骨和软骨组织替代治疗的自体 DPSCs 资源。
