In-vitro culture method of myoblasts derived from genioglossus of old mice

Abstract

The invention discloses an in-vitro culture method of myoblasts derived from genioglossus of old mice. The in-vitro culture method comprises the steps of placing the genioglossus of the old mice in aculture dish containing I-type collagenase, and cutting the genioglossus into pieces to obtain a mixed solution of genioglossus tissue fragments and I-type collagenase; placing the mixed solution in acentrifuge tube, and slightly blowing and beating to obtain a genioglossus suspension, performing primary digestion, centrifuging to remove supernatant, absorbing the I-type collagenase, and adding trypsin for secondary digestion; and centrifuging the genioglossus suspension obtained after digestion is ended, removing supernatant, re-suspending the obtained genioglossus fragments by using a primary culture medium, slightly blowing and beating, and uniformly mixing to prepare a suspension, then placing the suspension in an incubator, and culturing the suspension by adopting a differential adherent culture method and a semi-liquid-changing mode. According to the in-vitro culture method disclosed by the invention, the genioglossus myoblasts with high purity, high activity, high activity andextremely low pollution risk can be obtained, the cultured myoblasts are good in differentiation, high in cell activity, short in culture period and high in culture efficiency, and the in-vitro culture method has great popularization significance.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for primary isolation and culture of stem cells derived from the genioglossus muscle of aged C57BL / 6 mice. Background technique [0002] Obstructive sleep apnea-hypopnea syndrome (OSA) is a pathological disease prone to hypoventilation during sleep, and the incidence of OSA increases with age. The genioglossus is the main muscle that pulls the tongue forward in the upper airway dilator. It is called the "upper airway safety muscle". It plays a decisive role in the opening and size of the airway. The key to the pathway, its dysfunction is often closely related to the pathogenesis of OSA. Myoblasts play an important role in muscle growth and injury. When muscle fibers are damaged or stimulated, myoblasts begin to proliferate, differentiate and fuse to form new fibers and repair damaged muscle tissue. [0003] The myoblast culture model derived from aged ...

AI Technical Summary

Problems solved by technology

There are some methods for culturing genioglossus muscle myoblasts in the prior art, but these methods have two disadvantages: first, the genioglossus muscle is usually derived from young mouse genioglossus tissue, which cannot simulate the state of old age; second, the current At the stage, the mouse genioglossus myoblasts are often used for direct primary isolation, so the isolated cells are relatively low in purity and relatively poor in viability

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