Side population cells isolated from porcine dental pulp tissue with self-renewal and multipotency for dentinogenesis, chondrogenesis, adipogenesis, and neurogenesis
Side population cells isolated from porcine dental pulp tissue with self-renewal and multipotency for dentinogenesis, chondrogenesis, adipogenesis, and neurogenesis
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DOI：
10.1634/stemcells.2006-0161
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发表时间：
2006-11-01
期刊：
影响因子：
5.2
通讯作者：
Nakashima, Misako
中科院分区：
文献类型：
--
作者：
Iohara, Koichiro;Zheng, Li;Nakashima, Misako
作为对龋齿的再生反应，牙髓有可能形成牙本质。这种再生是由干/祖细胞介导的。因此，干细胞疗法可能在修复性牙本质的诱导中具有潜在的用途。我们通过流式细胞术排除 DNA 结合染料 Hoechst 33342，从牙髓中分离出侧群 (SP) 细胞，并将其自我更新能力和多能性与非 SP 细胞和原代牙髓细胞进行比较。 SP细胞的累积细胞数大于非SP细胞和原代牙髓细胞。 Bmi1在SP细胞中持续表达，表明SP细胞具有更长的增殖寿命和自我更新能力。接下来，研究了牙髓SP细胞的多系分化潜力的维持。 II 型胶原蛋白和聚集蛋白聚糖的表达证实了 SP 细胞的软骨形成转化（30%）。 SP细胞表达过氧化物酶体增殖物激活受体γ和接头蛋白2，显示脂肪形成转化。神经丝和神经调节蛋白的 mRNA 和蛋白的表达证实了神经源性转化（90%）。这些结果证明牙髓SP细胞保持多谱系分化潜力。我们进一步研究了骨形态发生蛋白2（BMP2）是否可以诱导牙髓SP细胞分化为成牙本质细胞。 BMP2 刺激三维颗粒培养物中牙本质唾液酸磷蛋白 (Dspp) 和釉质素的表达。将补充有 Bmp2 的 SP 细胞自体移植到断牙上可刺激修复性牙本质的形成。因此，成人牙髓含有 SP 细胞，这些细胞富含干细胞特性，可用于使用 BMP2 进行牙本质再生的细胞治疗。
Dental pulp has the potential to form dentin as a regenerative response to caries. This regeneration is mediated by stem/progenitor cells. Thus, stem cell therapy might be of potential utility in induction of reparative dentin. We isolated side population (SP) cells from dental pulp based on the exclusion of the DNA binding dye Hoechst 33342 by flow cytometry and compared its self-renewal capacities and multipotency with non-SP cells and primary pulp cells. The cumulative cell number of the SP cells was greater than the non-SP cells and primary pulp cells. Bmi1 was continuously expressed in SP cells, suggesting longer proliferative lifespan and self-renewal capacity of SP cells. Next, the maintenance of the multilineage differentiation potential of pulp SP cells was investigated. Expression of type II collagen and aggrecan confirmed chondrogenic conversion (30%) of SP cells. SP cells expressed peroxisome proliferator-activated receptor gamma and adaptor protein 2, showing adipogenic conversion. Expression of mRNA and proteins of neurofilament and neuromodulin confirmed neurogenic conversion (90%). These results demonstrate that pulp SP cells maintain multilineage differentiation potential. We further examined whether bone morphogenetic protein 2 (BMP2) could induce differentiation of pulp SP cells into odontoblasts. BMP2 stimulated the expression of dentin sialophosphoprotein ( Dspp) and enamelysin in three-dimensional pellet cultures. Autogenous transplantation of the Bmp2-supplemented SP cells on the amputated pulp stimulated the reparative dentin formation. Thus, adult pulp contains SP cells, which are enriched for stem cell properties and useful for cell therapy with BMP2 for dentin regeneration.