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Comparison of satellite cell-derived myoblasts and C2C12 differentiation in two- and three-dimensional cultures: changes in adhesion protein expression

机译:二维和三维培养中卫星细胞衍生的成肌细胞和C2C12分化的比较:粘附蛋白表达的变化

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Changes in the expression of adhesion proteins involved in myoblast differentiation were investigated in monolayer (two-dimensional) and 3D (three-dimensional) cell cultures. The expression of integrin alpha3 subunit, integrin beta1 subunit, ADAM12 (a disintegrin and metalloproteinase 12), tetraspanins CD9 and CD81 and M-cadherin were examined in the murine myoblast cell line C2C12 and in a primary culture of rat satellite cells. Myoblasts in monolayer and 3D cultures showed significant differences in their morphology and cytoskeletal organization. All of the studied proteins participated in myoblast fusion in each culture examined, but differences in their levels of expression were observed. Satellite cell-derived myoblasts exhibited higher expression of adhesion protein mRNAs than C2C12 cells. Also, C2C12 cells from a 3D culture showed slightly higher expression of adhesion protein transcripts than the same cells cultured as a monolayer. Significantly, the levels of adhesion protein mRNAs were found to change in parallel in all cell culture types. Despite this finding, it is important that differences between satellite cell-derived myoblasts and cell line C2C12 grown in monolayer and 3D cultures are taken into account when studying processes of myoblast differentiation in vitro.
机译:在单层(二维)和3D(三维)细胞培养物中研究了参与成肌细胞分化的粘附蛋白表达的变化。在鼠成肌细胞系C2C12和大鼠卫星细胞的原代培养物中,检查了整合素α3亚基,整合素β1亚基,ADAM12(整合素和金属蛋白酶12),四跨膜蛋白CD9和CD81和M-钙粘蛋白的表达。单层和3D培养物中的成肌细胞在形态和细胞骨架组织方面显示出显着差异。所有研究的蛋白质都参与了每种培养物中成肌细胞的融合,但是观察到它们表达水平的差异。卫星细胞衍生的成肌细胞比C2C12细胞表现出更高的粘附蛋白mRNA表达。同样,与以单层培养的相同细胞相比,来自3D培养的C2C12细胞显示的粘附蛋白转录物表达略高。值得注意的是,在所有细胞培养类型中,粘附蛋白mRNA的水平均发生平行变化。尽管有这一发现,但重要的是,在体外研究成肌细胞分化过程时,必须考虑到卫星细胞衍生的成肌细胞与单层和3D培养物中生长的细胞系C2C12之间的差异。

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