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Isolation and Biological Characterization of Muscle-Derived Stem Cells from Sheep Skeletal Muscle

Isolation and Biological Characterization of Muscle-Derived Stem Cells from Sheep Skeletal Muscle

 Ping Zhang1,2, Yabin Pu1, Yu Zhang2, Jia Chen2, Kunfu Wang3, Qian Li3, Yujiao Sun3, Yuehui Ma1, Shuqing Jiao2,* and Weijun Guan1,*

 1Institute of Beijing Animal Science and Veterinary, Chinese Academy of Agricultural Science, Beijing, 100194, China

2College of Pharmacy, Jiamusi University, Heilongjiang Province Key Laboratory of Biological Medicine Formulation, Jiamusi, 154007, Heilongjiang, China
3College of Wildlife Resources. Northeast Forestry University, No.26 Hexing Road, Xiangfang District, 150040, Harbin, China

Ping Zhang and Yabin Pu contributed equally to this work.

*      Corresponding authors: shuqingjiao60@163.com;  weijunguan301@gmail.com

 

ABSTRACT

The Objective of this study was to investigate the isolation, cultivation, identification and differentiation potential of Muscle-derived stem cells (MDSCs) from sheep skeletal muscle and provide an experimental evidence for clinical application. The skeletal muscle tissue were studied from fetal sheep, and the stepwise digestion by XI collagenase and trypsin was used to isolate MDSCs and were purified by differential attachment technique. Cell morphology were observed using an inverted microscope, and MDSCs proliferation pattern were determined through growth curve analyses. MDSCs were identified by immunofluorescence and RT-PCR, and the immunofluorescence antibody invoved Sca-1, CD34, CD144, Desmin and CD45 which are the makers of MDSCs. Finally, MDSCs were induced into Adipocytes, osteoblasts, chondrocytes and neuron-like cells by the optimized inducing medium. After the two-step digestion and differential attachment technique, high purity MDSCs were successfully cultured. MDSCs were identified by round or short spindle cell, and gathered into clusters; The average doubling time and clone efficiency of the cells were 35.5h and 47.2%, respectively. The MDSCs makers, such as Sca-1, CD34, CD144 and Desmin, were positive expressed by immunofluorescence and RT-PCR detection, and CD45 was negative expressed. MDSCs were successfully induced into Adipocytes, osteoblasts, chondrocytes, myoblasts and neuron-like cells under the optimized inducing medium. To conclude, in vitro high purity MDSCs have multiple differentiation potential, and can play an important role in muscle tissue repair, and provide seed cells for tissue engineering research and clinical applications.

 

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