Experimental Study On The United Culture Of Osteoblast And Bio-derived Bone In Vitro

PurposeBone marrow stroma cells ( BMSCs) is a kind of cell with potential differentiation ability, they can differentiate into osteoblasts under special condition, and can be obtained and expand easily.Bio - derived bone can be obtained from nature tissue by a serious of physical and chemistry method, so it is one of the best stand of tissue engineering.So we want to study cytobiological properties and osteogenic ability of osteoblast in vitro, which derive from BMSCs through separating, expanding, inducing, checking up ALP, and von kossa staining so on.Then united osteoblasts and bio - derived bone to cultivate biollgical active bone stand.Methods1 separate and cultivate BMSCs of rabbitAbout 4ml marrow were obtained from femurs after general anesthesia, put it into lymphocyte separate fluid of the same volume, centrifuged, got BMSCs, calculated number of cells, cultivated in the incubator. Invert phase - contrast microscopy was used to observe morphology change of BMSCs each day.2 Induce and differentiate BMSCs of rabbitThe BMSCs were induced to differentiate into osteoblasts by means of an induction fifferentiation medium containing dexamethasone, β - glycerophophate and vitamin C, then still use invert phase - contrast microscopy to observe mor-phology change of the cells each day.3 Identify osteoblasts induced from BMSCs3.1 examine ALP staining by Gomori calcium -cobalt staining3.2 examine the calcium node by von Kossa staining3. 3 examine the calcium node by aliarin - red staining4 Compound osteoblasts and bio - derived in vitro 4.1 prepare the bio - derived bone4. 2 compound osteoblasts and bio - derived by means of dipping and deposition5 Observe the result of compound culture through scanning electron microscopy6 StatisticsResult1 The morphologies of the first 3passages of BMSCs are stable, the BMSCs would get touch more than 80% in 7 ~ 10 days.2 BMSCs will differentiate into osteoblasts in 15days under special induction medium, and some special staining are positive.3 It can be seen that osteoblasts growing on the bio - derived bone in 7 days through scanning electron microscopy.Discussion1 The culture of BMSCsBecause of the anchoring feature of BMSCs, we isolated BMSCs from adult rabbit marrow by means of gradient centrifugation and anchoring culture. The BMSCs could be purified by some generations, this is a easy and effective method.2 The induction differentiation of osteoblastsALP is the function enzyme of osteoblast, it is a key standard of BMSCs differentiating into osteoblast the exercise results turned out that the number of ALP...