Effect of ischemic culture conditions on the survival and differentiation of porcine dental pulp-derived cells
Effect of ischemic culture conditions on the survival and differentiation of porcine dental pulp-derived cells
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DOI：
10.1111/j.1432-0436.2008.00282.x
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发表时间：
2008-11-01
期刊：
影响因子：
2.9
通讯作者：
Ueda, Minoru
中科院分区：
文献类型：
--
作者：
Agata, Hideki;Kagami, Hideaki;Ueda, Minoru
尽管分化和未分化的细胞在损伤或炎症的情况下可以暴露于缺血条件，但缺血对细胞存活和分化的影响尚未得到很好的表征。在这里，我们的特点猪牙髓衍生细胞（PDPCs）的培养条件下，近似缺血的反应。由于牙齿中狭窄的血管开口，牙髓经常暴露于缺血，这可能影响pDPC的分化状态。在这项研究中，我们研究了各种缺血条件对分化诱导和非诱导的pDPC的影响。为了了解本研究中所用细胞的特性，对已报道的牙髓干细胞的细胞表面标记进行了研究。pDPC为CD 90（低）、CD 105（+）和α-平滑肌肌动蛋白阳性，并显示成骨/软骨分化潜能。缺氧是最有害的因素，细胞活力，而缺氧并没有显着影响生存。葡萄糖浓度对细胞死亡具有显著的机制依赖性影响。葡萄糖的存在与半胱天冬酶依赖性细胞死亡相关，而葡萄糖的缺乏与半胱天冬酶非依赖性细胞死亡相关。相反，分化状态（即，诱导的与非诱导的pDPC）不影响细胞死亡的程度或机制。通过逆转录-聚合酶链反应在诱导和非诱导细胞中发现特异性标记物的耗竭表明细胞在缺氧下去分化。未诱导的pDPC对Oct-4、Sox-2和缺氧诱导因子-2 α的缺氧诱导敏感，而这些基因在诱导的pDPC中没有变化。再分化分析显示，从非诱导的pDPC的存活细胞显示两倍高的碱性磷酸酶活性相比，诱导的pDPC，这表明更大的可塑性之间的非诱导的pDPC的存活分数。这些数据表明，缺血条件对未分化和分化的pDPC具有相似的不利影响，并影响pDPC的分化状态。此外，缺血条件可能会影响未分化的pDPC的可塑性。
Although differentiated and undifferentiated cells can be exposed to ischemic conditions in cases of injury or inflammation, the effects of ischemia on cell survival and differentiation have not been well characterized. Here, we characterize the response of porcine dental pulp-derived cells (pDPCs) to culture conditions that approximate ischemia. Dental pulp is often exposed to ischemia due to narrow vascular openings in the tooth, which may affect the differentiation status of pDPCs. In this study, we investigated the influence of various ischemic conditions on differentiation-induced and non-induced pDPCs. To understand the character of cells used in this study, reported cell surface markers for dental pulp stem cells were investigated. pDPCs were CD90(low), CD105(+), and alpha-smooth muscle actin positive and showed osteogenic/chondrogenic differentiation potential. Anoxia was the most detrimental factor to cell viability, whereas hypoxia did not significantly affect survival. Glucose concentrations had a significant, mechanism-dependent effect on cell death. The presence of glucose correlated with caspase-dependent cell death, whereas the absence of glucose was linked to caspase-independent cell death. In contrast, differentiation status (i.e., induced versus non-induced pDPCs) did not affect the degree or mechanism of cell death. Finding depletion of specific markers by reverse transcription-polymerase chain reaction in both induced and non-induced cells suggests that the cells are de-differentiating under anoxia. Non-induced pDPCs were susceptible to anoxic induction of Oct-4, Sox-2, and hypoxia inducible factor-2 alpha, while these genes did not change in induced pDPCs. Re-differentiation analysis revealed that the surviving cells from non-induced pDPCs showed twofold higher alkaline phosphatase activity as compared with induced pDPCs, which suggest greater plasticity among the surviving fraction of non-induced pDPCs. These data showed that the ischemic conditions have similar detrimental influence on both undifferentiated and differentiated pDPCs, and affect differentiation status of pDPCs. Furthermore, ischemic conditions may influence the plasticity of undifferentiated pDPCs.