Culture Of Synovial Mesenchymal Stem Cells (SMSCs) From Tenosynovium Of Rabbit And Its Osteogenic Differentiation Potential In Vitro

Objective:Synovial membrane mesenchymal stem cell(SMSCs) is theimportant research topic of tissue engineering,SMSCs is acquired from the synoviallining layer of capsule of temporomandibular or knee jont, Another synovial membranesource,tenosynovium has the similar components, character of biochemistry andfunction，and the chance to become another source of SMSCs.We are trying to study themethod and feasibility of isolating the tenosynovium from rabbit achillestendon,culturing and obteining the purified SMSCs.further study the osteogenicdifferentiation potential in vitro,and discuss the possibility of application totendinopathy and osteoporosis.Methods:A3months old,1.5kilograms weight rabbit was killed by injecting airvia ear vein,with the help of surgery microscope, Synovial tissue was harvestedunder sterile conditions and microscopic magnification of the Achilles tendon sheath.The tissue was cut into small pieces,minced tissues were explant culture after digestingwith type II collagenase for one hour. Monoclone was obtained by selecting as primarySMSCs,under low planting density condition. The method of cytometry was used todraw a cellular growth curvethe at Passage3,4.Cell cycle analysis was performed byflow cytometry,Application of flow cytometry to detect the surface antigenCD44,CD45,CD90.The passage3cells were treated with inductive medium at,Cellmorphous was observed with an inverted microscope everyday.Osteogenicdifferentiation was tested by alkaline phosphatase and Von Kossa staining,separatelyon the tenth day,and3weeks later.Results:1.synovial tissue can be isolated from Achilles tendon sheath,after cultureand purification,the cells expanded steadyly and rapidly.Doubling time was42.5h.Surface antigen CD44and CD90wre positive,CD45is negative.After osteogenesis induction,the results for alkaline phosphatase and Von Kossa stainingwere postive.Conclusion:1.Synovial tissue is easy separated from Achilles tendon sheath,and SMSCs canbe isolated from it. Tenosynovium has the chance to take place of the joint synovium tobecome the new source of SMSCs,and is superior in tendinopathy.2.SMSCs from tenosynovium have the potential to differentiate into osteogenic invitro,and can be used for the therapy of osteoporosis...