Comparison of characteristics of periodontal ligament cells obtained from outgrowth and enzyme-digested culture methods
Comparison of characteristics of periodontal ligament cells obtained from outgrowth and enzyme-digested culture methods
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DOI：
10.1016/j.archoralbio.2010.10.013
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发表时间：
2011-04-01
影响因子：
3
通讯作者：
Izumi, Yuichi
中科院分区：
文献类型：
--
作者：
Tanaka, Keiko;Iwasaki, Kengo;Izumi, Yuichi
目的：牙周膜（PDL）细胞在牙周再生中发挥重要作用。已经研究了 PDL 细胞的独特特征，主要是源自 PDL 组织的生长过大的细胞。最近，利用酶消化从 PDL 组织中获得了间充质干细胞。通过两种方法（生长和酶消化）培养的 PDL 细胞特性的差异尚不清楚。本研究的目的是探讨通过这些方法获得的PDL细胞的特性。方法：采用生长法和酶消化法培养拔牙的PDL细胞。比较细胞增殖、集落形成活性以及向成骨细胞、脂肪细胞和软骨细胞的分化能力。还通过逆转录聚合酶链反应研究了 PDL 细胞、间充质干细胞和成纤维细胞的基因表达。使用酶联免疫吸附测定（ELISA）试剂盒测定I型原胶原c肽（PIP）的产生。结果：酶消化法培养的PDL细胞比生长法培养的PDL细胞表现出更高的增殖率、集落形成活性和向成骨细胞、脂肪细胞和软骨细胞的分化能力。 CD166（一种间充质干细胞标记物）在酶消化的 PDL 中略高于生长的 PDL，而 1 型胶原蛋白 α1 和 3 型胶原蛋白的基因表达在生长的 PDL 细胞中较高。此外，生长的PDL细胞比酶消化的PDL细胞表现出更高的PIP产量。结论：通过生长和酶消化获得的PDL细胞表现出不同的特征。酶消化法产生具有较高增殖率和间充质干细胞样特性的细胞，而在生长法中收集具有成纤维细胞样特性的细胞。不同培养方法的 PDL 细胞特性可能为发明 PDL 细胞的新治疗用途提供信息。 (C) 2010 Elsevier Ltd. 保留所有权利。
Objective: Periodontal ligament (PDL) cells have an important role in periodontal regeneration. The unique characteristics of PDL cells, mainly outgrown cells derived from PDL tissue, have been investigated. Recently, mesenchymal stem cells have been obtained from PDL tissue using enzyme digestion. The differences in properties of those PDL cells cultured by the two methods (outgrowth and enzyme digestion) are unclear. The objective of this study was to investigate the characteristics of PDL cells obtained by these methods.Methods: PDL cells from extracted tooth were cultured using outgrowth and enzyme digest methods. Cell proliferation, colony-forming activity and differentiation capacity to osteoblast, adipocyte and chondrocyte were compared. Gene expressions for PDL cells, mesenchymal stem cells and fibroblasts were also investigated by reverse transcription polymerase chain reaction. Procollagen type I c-peptide (PIP) production was measured using an enzyme-linked immunosorbent assay (ELISA) kit.Results: PDL cells cultured by enzyme digest methods showed a higher proliferation rate, colony-forming activity and differentiation capacity into osteoblast, adipocyte and chondrocyte than those in PDL cells by outgrowth method. CD166, one of the mesenchymal stem cell markers, was slightly higher in enzyme-digested PDL than in outgrowth PDL, whilst gene expressions for type 1 collagen alpha 1 and type 3 collagen were higher in outgrown PDL cells. Moreover, outgrowth PDL exhibited higher PIP production than enzyme-digested PDL cells.Conclusion: PDL cells obtained by outgrowth and enzyme digestion showed different characteristics. The enzyme digestion method yielded cells with higher proliferation rate and mesenchymal stem cell-like properties, whereas cells with fibroblast-like properties were collected in the outgrowth method. PDL cell properties by different culture methods may provide information for inventing new therapeutic uses of PDL cells. (C) 2010 Elsevier Ltd. All rights reserved.