牙乳头/牙囊细胞交互作用对细胞多能性的作用

摘要

AIM:To investigate the effect of cell-cell interaction of dental papilla cells (DPCs)and dental follicle cells (DFCs)on the pluripotency of the cells.METHODS:An in vitro cell-cell co-culture system of DPCs and DFCs was established.Cell growth,cell senescence and the expression of Oct-4,Sox2 and c-Myc were investiga-ted by cell counting,β-gal staining and immunohistochemical staining.The odontogenic differentiation capability of DPCs and DFCs was evaluated by ALP activity assay after 0,7,14,and 21 d of osteogenic induction.RESULTS:Under co-culture condition,cell growth of DPCs and DFCs was promoted,and cell senescence of DPCs and DFCs was inhibited.Expression of Oct-4,Sox2 and c-Myc was significantly elevated in both cells on day 5 after coculture.ALP activity was significantly upregulated in both cells after 14d and 21d of odontogenic induction.CONCLUSION:Opti-mized microenvironment with cell-cell communication may enhance the pluripotency potential of DPCs and DFCs.%目的：研究细胞交互作用对细胞多能性的调控作用。方法：建立牙乳头（DPCs）和牙囊细胞（DFCs）体外共培养模型，二者单独培养的细胞为对照组。采用细胞计数、β－gal 染色检测细胞生长和衰老情况；免疫荧光染色检测共培养条件下多能性相关因子 Oct－4、Sox2和 c－Myc 的表达变化；通过 ALP 活性测试检测矿化诱导后的 DPCs 和 DFCs 的矿化能力。结果：共培养组的 DPCs 和 DFCs 的细胞数明显高于对照组（P ＜0．05）；培养至第7代时，共培养组的 DPCs、DFCs 中与衰老相关的 SA－b－gal 表达较对照组明显减弱（P ＜0．05）；Oct－4、Sox2和 c－Myc 在共培养组 DPCs 和 DFCs 中的表达明显强于对照组；共培养组的 DPCs 和 DFCs 经矿化诱导14、21 d 后，其 ALP 活性均显著高于对照组（P ＜0．05）。结论：DPCs 和 DFCs 可通过体外交互作用抑制细胞衰老、促进细胞的多能性相关因子的表达、提高细胞的矿化能力。