| Objective Bone defect caused by wound> tumour and malformation is a familiar clinic disease. How to restore bone tissue and rebuild its dynam and biology system is one of the most important things of clinic researches. Thus the bone tissue engineering is making most development. With the development of bone tissue engineering, seeding cells are becoming one of the hotspots. At the present time, there are a lot of kinds of seeding cells. For example:bone,periosteum,marrow, tissue out of bone and forepart embryo, et al. All of them have their excellence and disadvantage. By now, the mesenchymal stem cells (BMSC) is the best one. Moreover, the tissue outside of bone, such as fat cells have great potential during the seeding cells. The text expatiate the reseaches about the isolating cultivate of processde lipoaspirate cells(PLA cells),and the differentiate into osteogenic lineage. The text wants to provide a science evidence for the choosing of seeding cells in clinical bone healing.Methods Adopting three-month old male Sprague-Dawley rat to put to death by the methed of dislocation. Isolating adipose-derived cells (ADSCs) from rat inguinal fat pads. Then digesed with Collagenase- I .After primary culture in DMEM culture fluid, I observed the growth and appearance of the cells by a inversion microscope. When the cells are becoming stabilizing, I separated them into 3 groups and incubated in induced medium of different density of DEX for 4 weeks to induce osteogenesis. Osteogenic differentiation was tested by alkaline Phosphatase staining. And detected by alkaline Phosphatase kit.Results (1) The primary culture and passage of adipose-derived stem cells(ADSCs).After primary cultivate, the cells are round or similar round. The cell body is lucency. floating in the culture fluid. There are a few of cells adhered after 24 hours. Afer 48 hours, the adherence cells are becoming more and more and begin to division growth.. The cells are growing by the style of disintegration colony. When the cells are tending to confluens, they are becoming slendemess. They are similar to mesenchymal stem cells(MSCs).(2) Osteogenesis induction and identify.After incubater in induced medium to induce osteogenesis, the cells are slowly changed. The shape of cells begin to transform into cube. This also similar to MSCs. After 4 weeks, the result for alkaline Phosphatase staining was positive. It indicate that there are expression of endogenous ALP. Von Kossa staining was positive indicate that the cells is osteoblast cells.(3) The assaying and comparision of alkaline Phosphatase.Separating the adipose-derived stem cells into 3 groups and incubated in induced medium of different density of DEX (10-9,10-8,10-7 mol/L for about 4 weeks .Then test the quantity of alkaline Phosphatase (ALP) by alkaline Phosphatase kit.All datas are analyzed by statistics software SPSS 12.0.With the ANOVA: P<0.05. Indicate that the dealing factors act to the result. Furthermore with the Multiple Comparisons: B vs A and C, P<0.05.But between A and C, P>0.05.Indicate that there is a difference between B and A or C. But the difference between A and C is not obviously.Collusions Adipose-derived stromal cells(ADSCs) can be isolated from rat adipose tissue after osteogenesis induction. Their biological characteristics are analogy to mesenchymal stromal cells(MSCs),and have the potential to differentiate into osteogenic lineage. The expression of alkaline Phosphatase is higher in the duced medium withing the dexameth density of 10-8 mol/L.
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