The ensured proliferative capacity of myoblast in serum-reduced conditions with Methyl-β-cyclodextrin

by Selleckchem | Oct 04 2023

To produce muscle fibers for cultured meat on a large scale, it is important to expand myoblasts in a serum-reduced or serum-free medium to avoid cost, ethical, and environmental issues. Myoblasts such as C2C12 cells differentiate quickly into myotubes and lose their ability to proliferate when the serum-rich medium is replaced with a serum-reduced medium. This study demonstrates that Methyl-β-cyclodextrin (MβCD), a starch-derived agent that depletes cholesterol, can inhibit further differentiation of myoblasts at the MyoD-positive stage by reducing plasma membrane cholesterol on C2C12 cells and primary cultured chick muscle cells. Furthermore, MβCD efficiently blocks cholesterol-dependent apoptotic cell death of myoblasts, which is one of the mechanisms by which it inhibits the differentiation of C2C12 myoblast cells, as dead cells of myoblast are necessary for the fusion of adjacent myoblasts during the differentiation process into myotubes. Importantly, MβCD maintains the proliferative capacity of myoblasts only under differentiation conditions with a serum-reduced medium, suggesting that its mitogenic effect is due to its inhibitory effect on myoblast differentiation into myotube. In conclusion, this study provides significant insights into ensuring the proliferative capacity of myoblasts in a future serum-free condition for cultured meat production.

Comments:

The passage you provided discusses a study that explores the use of Methyl-β-cyclodextrin (MβCD), a compound derived from starch, to inhibit the differentiation of myoblasts into myotubes. The aim of the study is to maintain the proliferative capacity of myoblasts, which is crucial for producing muscle fibers for cultured meat on a large scale.

In traditional cell culture techniques, myoblasts are typically cultured in a serum-rich medium that promotes their differentiation into myotubes. However, using serum-rich media can be costly, raise ethical concerns (as it often involves the use of animal-derived serum), and have environmental implications. Therefore, finding alternative methods to sustain myoblast proliferation in a serum-reduced or serum-free medium is important.

The study demonstrates that MβCD can inhibit further differentiation of myoblasts at the MyoD-positive stage. MyoD is a protein involved in the regulation of myoblast differentiation. By reducing plasma membrane cholesterol on C2C12 cells (a mouse myoblast cell line) and primary cultured chick muscle cells, MβCD prevents myoblasts from progressing into the myotube stage.

Additionally, MβCD is shown to block cholesterol-dependent apoptotic cell death of myoblasts. This mechanism contributes to its inhibitory effect on myoblast differentiation. It is important because the presence of dead myoblasts is necessary for the fusion of adjacent myoblasts during the differentiation process into myotubes.

Crucially, MβCD only maintains the proliferative capacity of myoblasts under differentiation conditions with a serum-reduced medium, suggesting that its mitogenic effect is specifically due to its inhibitory effect on myoblast differentiation into myotubes.

In summary, this study provides valuable insights into maintaining the proliferative capacity of myoblasts in a future serum-free condition, which is essential for the large-scale production of muscle fibers for cultured meat. By using MβCD to inhibit myoblast differentiation and apoptosis, researchers can potentially overcome the limitations associated with serum-rich media and contribute to the development of sustainable and ethical cultured meat production methods.