Effect of Gelatin on Osteogenic Cell Sheet Formation Using Canine Adipose-Derived Mesenchymal Stem Cells
Effect of Gelatin on Osteogenic Cell Sheet Formation Using Canine Adipose-Derived Mesenchymal Stem Cells
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DOI：
10.3727/096368916x693338
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发表时间：
2017-01-01
影响因子：
3.3
通讯作者：
Kweon, Oh-Kyeong
中科院分区：
文献类型：
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作者：
Kim, Ah Young;Kim, Yongsun;Kweon, Oh-Kyeong
使用间充质干细胞（MSC）的成骨分化细胞片技术可用于刺激骨再生。细胞片层技术的优点是将活细胞有效地递送到聚焦区域中。我们开发了一种新的成骨细胞片技术，通过添加明胶成骨细胞培养基。明胶诱导的成骨细胞片（GCS）进行了比较，传统的成骨细胞片（OCS）。未分化的MSC（UC）用作对照。在显微镜下和组织学上评价这些细胞片的形态。通过DNA定量估计时间依赖性细胞增殖率。成骨基因标志物的表达和钙沉积的数量分别通过定量实时聚合酶链反应和茜素红S（ARS）染色进行评估。GCS比OCS更厚更强。与OCS相比，GCS显示出显著更高的细胞增殖率（p < 0.05）。与OCS相比，GCS表现出显著更高的BMP-7 mRNA上调（p < 0.05）。GCS和OCS在第10天均显示ARS反应阴性，但仅GCS在第21天显示ARS反应阳性。通过这种技术，我们观察到活跃的细胞增殖与丰富的ECM和成骨骨标志物的上调，我们的研究结果表明，GCSs可能是有前途的骨再生的治疗应用。
Osteogenically differentiated cell sheet techniques using mesenchymal stem cells (MSCs) are available to stimulate bone regeneration. The advantage of the cell sheet technique is delivering live cells effectively into the focal region. We developed a novel osteogenic cell sheet technique by adding gelatin to osteogenic cell medium. Gelatin-induced osteogenic cell sheets (GCSs) were compared to conventional osteogenic cell sheets (OCSs). Undifferentiated MSCs (UCs) were used as a control. The morphology of these cell sheets was evaluated microscopically and histologically. The time-dependent cell proliferation rate was estimated by DNA quantification. The expression of osteogenic gene markers and the number of calcium depositions were assessed by quantitative real-time polymerase chain reaction and Alizarin red S (ARS) staining, respectively. GCSs were thicker and stronger than OCSs. GCSs showed a significantly higher cell proliferation rate compared to OCSs (p < 0.05). GCSs exhibited significantly higher upregulation of BMP-7 mRNA compared to OCSs (p < 0.05). Both GCSs and OCSs showed negative ARS reactivity on day 10, but only GCSs showed positive ARS reactivity on day 21. With this technique, we observed active cell proliferation with abundant ECM and upregulation of osteogenic bone markers, and our results suggest that GCSs could be promising for therapeutic applications in bone regeneration.