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| Isolation, Culture and Identification of Dairy Goat (Capra hircus) Myoblast In vitro |
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Abstract Myoblast is capable of self-renewal and polymorphism, and plays an important role in the regeneration and repair of skeletal muscle. In order to establish a method for isolation and purification, culture and identification of dairy goat (Capra hircus) myoblasts in vitro, and to understand the identification of myoblast features of proliferation and myogenesis, and to provide experimental materials for the regulation mechanism of muscle development in dairy goats, the samples were taken from skeletal muscle of Xinong Saanen dairy goat kids. Cells were disassociated with collagenase I and trypsin following with purification of cell using differential adhesion approach. qRT-PCR and immunofluorescence staining were conducted to further identify purified cells. Cell morphological results showed that purified cells were in spindle shape and maintained a stable status of growth. Myogenic regulatory factors, myogenic differentiation factor (MyoD) and myogenic regulatory factor 5 (Myf5), and desmin were positively expressed in purified cells (percentage of purified cell ＞ 95%) as observed from the results of qRT-PCR and immunofluorescence staining. Purified myoblasts were able to fuse into myotubes after induced differentiation. Myosin heavy chain (MYHC), a specific marker of differentiated myotubes, was positively expressed. This study establish a method of isolation and culture of dairy goat myoblasts in vitro. Myoblast line with high purity is successfully obtained, which is able to be used as an experimental material in studies of regulation mechanism of muscle development in dairy goats.
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Received: 24 April 2018
Published: 24 October 2018
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