| The common muscle injury of athletes is mainly divided into three kinds of muscle tissue sprain,pull and contusion.Muscle injury has a high incidence in sprint,and muscle injury is the first injury of sprinters,which seriously affects athletes' functional level,athletic ability and athletic performance.Research on repair and treatment of muscle injury has become a popular trend in clinical medicine and sports medicine.At this stage,there are many ways to repair damaged muscles,such as drug therapy,surgical treatment,acupuncture and massage,physical therapy,etc.After clinical verification,the above treatment has a certain effect on muscle injury,but the treatment cycle is long,it is easy to produce scar and reduce muscle function.Stem cells in human growth and development and plays a crucial role in the regeneration and repair of damaged tissue,during the national “12th Five-Year” stem cells,the treatment of sports injury by leaps and bounds,fruitful,“13th Five-Year” will be included in the planning of stem cell research,sports and medical field of stem cell therapy in the future full of confidence.The purpose of this study was to isolate skeletal muscle stem cells in vitro,establish the amplification culture system of muscle satellite cells,and detect the proliferation and differentiation of biological characteristics of skeletal muscle satellite cells,muscle injury animal model design,injection of muscle satellite cells were detected after intervention,transplanted stem cell migration and repair in vivo,the experimental results are as follows:1.By mechanical separation,the separation of fetal sheep anterior tibial muscle source of type I collagenase and trypsin digestion method of muscle satellite stem cells,the cells obtained by purified and cultured to the 31 generation differential sorting method,after cryopreservation and resuscitation rate of living cells still have high specific surface markers.Identification of on the in vitro proliferation of stem cells by RT-PCR,flow cytometry and immunofluorescence results showed that the cells expressed a higher positive rate of MYOG,MYF5,PCNA,C-MET and desmin genes.The results show that the characteristic after screening of cells with muscle satellite cells.2.The “stemness” study of the cultured cells was carried out.According to the characteristics of cell expansion detection cloning ability,growth curve.Culture medium containing different factor induce cells into adipocytes,osteoblasts,chondrocytes,myoblasts differentiation,differentiation of the cells was detected by RT-PCR immunohistochemical technique and chemical related gene expression,the results were positive.It shows that the isolated cells conform to the characteristics of stem cells and are muscle satellite cells.3.A mouse muscle injury model was established by Cytotoxin(CTX).Paraffin wax sections were stained to observe that most of the normal muscle tissues were damaged.Serological detection showed that the creatine kinase(CK)in the blood of mice was significantly higher than the normal range.The muscle satellite stem cells labeled by CM-Dil cell tracer were injected into the muscle injury site of mice directly,and 7 time gradients were set.Each gradient was taken for paraffin sections,frozen sections and serological tests.The results showed that the transplanted stem cells had a good repair effect on the muscle damage.In summary,this study analyzed the sprinters muscle injury and stem cell research,successfully isolated muscle satellite cells in vitro,and the “stemness” verified by muscle satellite cells to repair muscle stem cell injury in mice,and provide a theoretical basis for promoting tissue regeneration,and provide experimental basis for clinical application of stem cells the treatment of sports injury. |
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