Proliferative activity of skeletal myoblast sheet by paracrine effects of mesenchymal stem cells
Proliferative activity of skeletal myoblast sheet by paracrine effects of mesenchymal stem cells
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DOI：
10.1016/j.job.2016.05.005
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发表时间：
2016-11-01
影响因子：
2.4
通讯作者：
Abe, Shinichi
中科院分区：
文献类型：
--
作者：
Umezawa, Takashi;Higa, Kazunari;Abe, Shinichi
目的：探讨骨髓间充质干细胞(MSC)对骨骼肌成肌细胞增殖活性的促进作用。方法：用IA型胶原凝胶(MC+)与兔骨骼肌成肌细胞片(MC+)和不含MC-细胞片(MC-)人工构建成肌片。结果：苏木精-伊红染色结果显示，第7天MC+较MC-厚，免疫组织化学染色显示，第2天两片结蛋白表达较低，第7、12天结蛋白表达增加，脱蛋白定位与层状区一致。逆转录-聚合酶链式反应检测到第2、7、12天两张肌片均有MyoD和PAX7的表达，第7天和第12天两张均可见MyoD和PAX7的表达。肝细胞生长因子在第2天MC+和第7天MC+均有明显表达。DNA细胞周期分析显示第7天MC+的细胞增殖活性明显高于MC-。第12天MC+和MC-的细胞增殖活性无显著差异。结论：MSCs的存在可一过性增强成肌细胞的增殖活性。但由于接触抑制，这种增强最终会减弱。(C)2016年日本口腔生物学协会。爱思唯尔出版，版权所有。
Objectives: The purpose of this study was to examine the proliferative activity enhancement of skeletal myoblasts in the presence and absence of mesenchymal stem cells (MSC).Methods: We artificially fabricated two types of cell sheets by co-culturing rabbit skeletal myoblast sheets with rabbit MSCs using type IA collagen gel (MC+), and cell sheets without rabbit MSCs (MC-).Results: The results of hematoxylin and eosin staining revealed that MC+ was thicker than MC- on day 7. Immunohistochemical staining revealed a low level of desmin expression in both sheets on day 2. Desmin expression increased at days 7 and 12, and desmin localization was consistent with the stratified area. Reverse transcription PCR revealed the presence of MyoD and PAX7 in both sheets on days 2, 7, and 12. The presence of myogenin was confirmed in both sheets on days 7 and 12. Hepatocyte growth factor expression was evident in MC+ on day 2, and in both sheets on day 7. Measurement of cell proliferative activity based on DNA cell cycle analysis indicated that MC+ had significantly higher cell proliferative activity than MC- on day 7. There were no significant differences in cell proliferative activity between MC+ and MC- on day 12.Conclusions: This study demonstrated that the presence of MSCs could transiently enhance the proliferative activity of myoblasts, but that this enhancement is ultimately diminished due to contact inhibition. (C) 2016 Japanese Association for Oral Biology. Published by Elsevier B.V. All rights reserved.