绵羊骨骼肌卫星细胞分离培养、鉴定与成肌诱导分化

doi:10.16431/j.cnki.1671-7236.2021.04.007

Abstract

Abstract: This experiment was aimed to isolate sheep skeletal muscle satellite cells(SMSCs),establish an optimal system for isolation,culture and identification of sheep SMSCs in vitro,thereby laying a seed cells for future pertinent researches.The SMSCs were isolated from new-born healthy sheep,disassociated with collagenase Ⅳ and trypsin and purified via differential adhesion method.RT-PCR and immunofluorescence methods were used to identify the expression of SMSCs marker genes paired box 7 (Pax7),Desmin and myogenic regulatory factors 1(MyoD1),and serum withdrawal method was used to induce SMSCs to differentiate into myoblasts.Satellite cells underwent muscle differentiation and formed myotubes.Muscle differentiation marker myosin heavy chain (MHC) in cytoplasm were detected by immunofluorescence.RT-PCR results showed that the amplified bands were consistent with the expected results,indicating that the cells expressed SMSCs marker genes Pax7,Desmin and MyoD1.The cultured cells expressed marker proteins Pax7,Desmin and MyoD1,demonstrating that SMSCs were obtained.Muscle differentiation marker MHC was detected in these cells.From the above results,the isolated and cultured cells were SMSCs,and a suitable in vitro culture system for sheep SMSCs was established,and myogenic differentiation was successfully carried out,which provided experimental materials and technical support for the future research on the mechanism of sheep skeletal muscle growth and development.

Key words: sheep; skeletal muscle satellite cells; isolation and culture; immunofluorescence identification; myogenic differentiation

CLC Number:

S826

LI Xin, YU Yongsheng, ZHANG Lichun, MA Huihai, LUO Xiaotong, WEI Tian, XIAO Cheng, ZHANG Qi, CAO Yang, ZHAO Zhongli. Isolation,Culture,Identification and Myogenic Differentiation of Sheep Skeletal Muscle Satellite Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(4): 1204-1210.

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Isolation,Culture,Identification and Myogenic Differentiation of Sheep Skeletal Muscle Satellite Cells

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