Broiler Amniotic Mesenchymal Stem Cells For Treatment Of Collagenase Ⅰ-induced Mouse Tendon Injury In Animal Models

Objective:Injury and illness has always been a major problem affecting the athletic performance and career of athletes.Among them,tendon injuries are the most common sports diseases in sports competitions,resulting in abnormal performance in competitive level and athletic ability,and they have become one of the urgent problems to be resolved in sports medicine.Stem cells can differentiate into multiple germ layers and play a significant role in the repair of damaged tissues.Methods:In this study,two different types of amniotic mesenchymal stem cells(AMSCs)were cultured in vitro and subcultured to a certain system to detect the biological characteristics of AMSCs and compare the difference of biological characteristics between the two stem cells derived from different sources,in which way,cells with better quality were chosen to serve research basis for tendon treatment.Animal injury model was established and the therapeutic effect of stem cells on tendon repair was explored in depth,arriving at the following experimental results:Results:1.Use type II collagenase and trypsin for co-digestion of amniotic tissue of broiler chicken which were then inoculated in six-hole plates after isolation and purification and cultured in vitro to 22 generations.RT-PCR and immunofluorescence were used to detect the expression of stem cell surface markers derived from amniotic tissue.Immunofluorescence results showed that the genes with positive expression of cells were:CD44,CD73,and CD90;RT-PCR results suggested that:CD29,CD44,and CD90 were expressed with no expression in CD34.The biological characteristics of the cells were obtained,and the cells were induced to differentiate into adipogenic,osteogenic,chondrogenic,and tendon cells.RT-PCR test indicated that specific genes were all positive.2.Establish mouse tendon injury model and damage mouse tendon with local injection of type I collagenase.Observe the injury condition of the tendon tissue using pathological section of paraffin and detect the change of creatine kinase(CK)content in the blood of mouse.AMSCs were inoculated onto the injured part of the mouse after being labeled with fluorescence and concentration of cells at the injured site were detected by applying frozen section.Tissue sections and serological analysis suggested that the structure and function of tendons were well repaired.Conclusions:This study will focus on the biological characteristics of AMSCs and their important role in tendon repair,laying theoretical foundations for stem cell therapy and providing experimental basis for subsequent clinical application.