Biological Characterization Of Bone Marrow Stromal Cell Of Rabbits Cultured In Vitro

Objective:Adult human stem cells that are intrinsic to various tissues have been described and characterized recently.These cells are capable of maintaining,generating, and replacing terminally differentiated cells within their own specific tissue as a consequence of physiologic cell turnover or tissue damage due to injury.Recent data suggest that adult stem cells generate differentiated cells beyond their own tissue boundaries,a process termed" developmental plasticity." In this article we focus on in vitro models of adult stem cells derived from bone marrow and their potential therapeutic applications.Stem cells are defined as cells that have clonogenic and self-renewing capabilities and that differentiate into multiple cell lineages.It is demonstrated that bone stromal stem cells(BMSCs)have multiple potential to differentiate into muscle,bone, cartilage,adipose,blood vessel endothelial,liver and neuron cells.BMSCs can be used as cell source in tissue engineering,cell implant and gene therapy with the possibility to be distributed to different tissue and organs normal circulation.They hold great promise in future therapeutic application in that they are easy to obtain,separate,expand and transfect,the procedure of bone marrow extraction is less harmful and the replantation induces no immune rejection.Our purpose in this study is to to establish a simple and stable method to culture the bone marrow stromal cells(BMSCs)from rats and reduce them to osteoblasts in vitro,and explore the correlation between proliferation and differentiation of BMSCs cultured in vitro to provide seed cells for bone tissue engineeringMethods:The bone marrow was separated from the bone of limbs of rats and was isolated and purified using adherence filtration.The passage cells of the third generation were divided into groups:1.BMSCs were cultured for 20days in the common medium and in the conditional medium,changed the liquor once 3 days.Draw the growth cure of each.2.Culured in the conditional medium(adding 10^-7mmol/L desamethasone,10mmol/Lβ-glycerol phosphate and 50mg/L Vitamin C in DMEM),, changed the liquor once every 3 days in the control group and in the test group,passaged once every 4 days in the test group,ALP(Alkaline phodphatase)was measured in each group once every 4 days.The formation of calcium nod was observed with Von Kossa' staining in the 20^thday.Result:1.The growth velocity of the cells cultured in the common medium was obviously faster than that in the conditional medium.2.There was a signicicant difference of the amount of ALP between the control group and the test group(P＜0.05),the amount of the test group is lower than that in the control group.The Von Kossa' staining in the test group was negative in the test group and positive.Conclusion:1.Adhering and screening method is a easy and reliable method to separate BMSCs.2.BMSCs can be obtained easily and have a strong osteogenic capacity,so BMSCs can be served as the seeding cells in the bone tissue engineering.3.There is a mutual depressive relationship between the differentiation and proliferation of BMSCs.