鹅成肌细胞的体外培养、鉴定及温度对细胞增殖分化的影响

In vitro culture and identification of Magang goose myoblasts and effect of temperature on cell proliferation and differentiation

骨骼肌的生长发育与动物生长性状有很强的关联性,建立鹅成肌细胞培养方法对于鹅骨骼肌生长发育的研究具有重要意义.本研究使用马岗鹅胚胎期15 d左右的胸腿肌组织,通过胰蛋白酶消化法分离细胞,差速贴壁法纯化成肌细胞;通过对成肌细胞分化决定因子Myog和成熟肌管标志基因Myh1的表达量进行检测,并结合肌管标志蛋白MyHC免疫荧光染色对分离纯化并诱导分化的成肌细胞进行鉴定.本研究发现腿肌成肌细胞增殖分化能力显著高于胸肌,成肌细胞在39℃培养温度下增殖分化能力高于37℃.本研究成功建立了马岗鹅胚成肌细胞系,对后续进行鹅骨骼肌发育调控机理研究具有重要意义.

Skeletal muscle development is strongly correlated with growth traits and establishing methods of goose myoblast isolation in vitro is vital for goose muscle development studies.Chest/leg muscle from Magang goose E15 was isolated by trypsin digestion and purified by differential attachment.Myogenic cells were identified by myogenic differentiation marker Myog and mature myofiber marker Myh1,together with immunofluorescence staining of myofiber marker MyHC.It was found that leg muscle oriented myogenic cells showed significantly higher proliferation and differentiation ability when compared to chest muscle,and myogenic cells under 39℃culture showed higher proliferation and differentiation ability compared to that under 37℃.The method of goose embryonic myoblast isolation in vitro,which would benefit goose muscle development studies was successfully established.