Competence of In Vitro Cultured Mouse Embryonic Stem Cells for Myogenic Differentiation and Fusion with Myoblasts
Competence of In Vitro Cultured Mouse Embryonic Stem Cells for Myogenic Differentiation and Fusion with Myoblasts
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DOI：
10.1089/scd.2013.0582
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发表时间：
2014-10-15
影响因子：
4
通讯作者：
Moraczewski, Jerzy
中科院分区：
文献类型：
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作者：
Archacka, Karolina;Denkis, Agnieszka;Moraczewski, Jerzy
多能干细胞是用于再生医学的各种细胞类型的潜在来源。尽管积累了知识，但目前还没有不需要遗传操作的有效且可重复的方案来从多能干细胞产生肌原细胞。在这里，我们研究了小鼠胚胎干（ES）细胞是否能够进行肌分化和融合的信号释放分化成肌细胞。使用表达组蛋白2B-绿色荧光融合蛋白的ES细胞，我们能够检测由ES细胞和分化的成肌细胞形成的杂交肌管。与成肌细胞融合的ES细胞下调多能性标记物的表达并诱导肌源性标记物的表达，而未融合的ES细胞没有表现出这种表达模式。因此，成肌细胞释放的信号不足以诱导ES细胞的肌分化。虽然ES细胞合成许多蛋白质参与成肌细胞粘附和融合，我们没有观察到任何肌管形成专门由ES细胞。我们发现，ES细胞缺乏M-钙粘蛋白和血管细胞粘附分子-1，这可能是解释低频率的混合肌管形成在ES细胞成肌细胞共培养和ES细胞无法单独形成肌管。
Pluripotent stem cells are a potential source of various cell types for use in regenerative medicine. Despite accumulating knowledge, there is currently no efficient and reproducible protocol that does not require genetic manipulation for generation of myogenic cells from pluripotent stem cells. Here, we examined whether mouse embryonic stem (ES) cells are able to undergo myogenic differentiation and fusion in response to signals released by differentiating myoblasts. Using ES cells expressing the histone 2B-green fluorescent fusion protein, we were able to detect hybrid myotubes formed by ES cells and differentiating myoblasts. ES cells that fused with myoblasts downregulated the expression of pluripotency markers and induced the expression of myogenic markers, while unfused ES cells did not exhibit this expression pattern. Thus, the signals released by myoblasts were not sufficient to induce myogenic differentiation of ES cells. Although ES cells synthesize many proteins involved in myoblast adhesion and fusion, we did not observe any myotubes formed exclusively by ES cells. We found that ES cells lacked M-cadherin and vascular cell adhesion molecule-1, which may account for the low frequency of hybrid myotube formation in ES cell-myoblast co-cultures and the inability of ES cells alone to form myotubes.