Subculture purification method of sheep embryo-derived myoblasts
A technology for myoblasts and purification methods, applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., to achieve the effects of enriching cell populations, high gene retention, and stable proliferation levels
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Embodiment 1
Myotubes are special structures formed by highly differentiated myoblasts. Myotubes, also known as multinucleated muscle fibers, are formed by the contact and fusion of multiple myoblasts. Microscopic observation results and morphological characteristics show that X10 myoblasts have obvious myotube structure different from other generations ( figure 2 ), combined with the analysis of fluorescence quantitative results, the X10 myoblast MYF5 Gene expression was significantly higher than other generations including X1 ( image 3 ). show MYF5 Gene expression is closely related to myotube formation. During the proliferation and differentiation of myoblasts, the expression of myoblast marker genes is down-regulated with the increase of generations, and once myotubes are formed MYF5 Gene expression will be significantly up-regulated without being affected by increasing generations. In addition, the above five genes are closely related to the proliferation and differentiation ...
Embodiment 2
[0054] Myoblasts are the precursor cells of skeletal muscle and are muscle-derived stem cells derived from the mesoderm. They have the ability of self-renewal and muscle fiber regeneration. Myoblasts mainly exist in embryonic and mature muscle tissues. In embryos, myoblasts undergo After proliferation and differentiation, multinucleated muscle fibers, namely myotubes, are fused to form a hollow tubular structure formed by the fusion of myotube cells. The nuclei move to the edge of the myotube and are difficult to be observed. With cell polarity, irregular distribution, no direction.
[0055] The result of this comparative example shows: the ratio of myoblasts in the cells purified by the conventional purification method is relatively low, and it is difficult to voluntarily differentiate to form myoblast-specific myotube structure; Differentiate to form myoblast-specific myotube structures.
[0056] Although the present invention has been described in detail with general descr...
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