Detection Of Biological Characteristics Of Muscle Satellite Cells And Tendon Stem Cells And Their Study In Animal Models Of Muscle And Tendon Injury

Sports injuries mainly referred to a variety of injuries occurred during exercise,which is often the site of injury and the technical characteristics related to the movement, including the main muscle tissue strain, sprains and contusions, as well as tendon and ligament injury. These injuries not only affect their health and impede their daily life and work, that currently appears in the medical field of stem cell therapy treatment of related diseases such high hopes. This study aimed to establish in vitro skeletal muscle satellite cells and stem cells were cultured tendon system, and explore its cell proliferation, differentiation and other biological characteristics and in the muscle and tendon injury injury model in vivo transplantation model migration traits, get the following conclusions:1. Mechanical separation and enzymatic digestion, successfully isolated muscle satellite cells and stem cells in tendons,strong cell proliferation after two subcultures separated,wherein muscle satellite cells spread to 27 generations,tendon stem cells spread to 38 generations, and freeze after the deposit muscle satellite cells has a higher cell activity.2. By RT-PCR, immunofluorescence and flow cytometry for cell-associated specific genes and surface markers were detected, showed muscle satellite cells Desmin,Pax7,c-Met,Myf5,Myo D,tendon stem cells in collagen I,collagen and antigen-specific gene expression showed III, CD44, Nucleostemin Tenascin-C and other surfaces, indicating that the cells were isolated stem cells have characteristic and tendons muscle satellite cells, in addition to the separation of the two cells growth curve and colony formation biological characterization capability,karyotype analysis, indicates that the cell has a strong ability to add value and no chromosomal mutation, in order to ensure the present and future experiments get use to build two research stem cells can be used for cell libraries.3. To identify muscle satellite cells more differentiation potential properties, in vitro muscle satellite cells, respectively, to myoblasts, into adipocytes and osteoblasts was induced, and the tendon of stem cells is to differentiate intoadipocytes, osteoblasts and chondrocytes inducing and induced results were identified, the results showed that: muscle satellite cells were significantly changed in the 7 days after the myoblasts induce morphological,by immunohistochemistry of fast skeletal myosin antibody staining was positive,RT-PCR to detect muscle-related genes MyoG and fast skeletal myosin were positive expression; tendon stem cells into chondrocytes induced cartilage ball shape obvious, alcian blue stained positive,RT-PCR related genes Collagen II, SOX9 and ACAN were expressed; in addition to the two kinds of cells in adipogenic and They are obtained when osteogenic success using liquid fat induced muscle satellite cells adipogenic, 21 days after oil red O staining can be seen in red lipid droplets, RT-PCR detected adipocyte-specific molecular marker LPL and PPAR-? expression; muscle satellite cells induce osteogenic medium after 4 weeks alizarin red staining was positive, RT-PCR detected osteoblast-specific molecular markers Collagen I, OPN, RunX2 and ALP expression; research shows that more than muscle satellite cells with multiple differentiation potential characteristics in the process of in vitro cultured.4. Respectively cardiac toxin(CTX) and collagenase build muscle damage in mice model and tendon damage model, H & E staining showed normal muscle and tendon tissue structure were destroyed, serology find CK indicators are significantly beyond the normal range indicate successful modeling. Muscle satellite cells were labeled tracer CM-Dil cells, the cells were labeled directly injected into mice muscle injury site point, fluorescence microscopy cells, serum-containing tissue to detect visible marks show stem cell transplantation CK value and damage and saline groups decreased compared, the results show that the two models of stem cell transplantation stable migrate into the muscle tissue in three weeks with a muscle injury in mice some recovery action.In summary, this experiment is successful in vitro culture of isolated muscle satellite cells and tendon stem cells, and biological characteristics were studied and multi-differentiation potential, in vitro mouse model transplanted muscle damage,muscle satellite cell transplantation in three weeks of steady migration to the appropriate tissues, muscle damage in mice with a certain recovery effect, and initially to stem cell-based therapies for treating damaged cells to promote tissueregeneration and repair of the clinical application to provide experimental basis.