| objectiveThe research of the stem cells has important significance to human being's health. There are a lot of problems on the clinic using of tissue-engineering research and one of them is the resource of seed cell. It was the excellence of pluripotentiality differentiation and strong ability of proliferation that made bone marrow stromal cells (BMSCs) becoming the main seed cell on tissue-engineering research.In this research, BMSCs was isolated and proliferation in order to explore an appropriate condition of isolation and culture system in vitro. At the same time,to found a technique and theory method on the clinic application.. The research on BMSCs also has the significance on new recognizing of cell isolation,proliferation and differentiation. In this research,the effects on proliferation and differentiation of BMSCs being cultured by Bushen herbs-content serum was anlysised. Develop the benefits of traditional Chinese medicine and connect them with the stem cells research so they can promote each other,and explore a way for the research on basic theory of stem cell in Traditional Chinese Medicine characteristic. In the same time,it may supply support on theory in microworld to detect the mechanism of Bushen herbs and application on clinic of Bushen method.By exploring the methods of the herbs-content serum preparation and observing the effects of single Bushen drug on proliferation and differentiation of BMSCs,the theropy mechanism of the Bushen herbs on ordinary disease on bone department was discussed.MethodsAfter being isolated and cultured in vitro,the growth character and the growth curvewas of BMSCs were obtained by morphological observition and MTT colorimetry respectively.In the process of the single Bushen-herb serum preparation,it is important in the selecting of animal,dosage and project of gastric infusion etc. Analysis the effects of the different preparation methods of drug-content serum on serum quality and getting an optimization methods of drug -content serum.The effects on proliferation of Bushen herbon BMSCs was observed with the number of culture days of F2 BMSCs along abscissa versus the number of the cells along vertical axis while none drug-content serum being the negative control. the BMSCs were cultured in Bushen herb-content serum and the osteogenesis inducer was positive control, after two generations,the effects of the Bushen herbs-content serum on BMSCs' differentiation were observed.ResultBMCSs have strong ability of adapting in vitro,by using adherence separation method,cells can be finished primary culture process in 10-12 days.F2 generation grew rapidly,passaged by 1:3 and after 4 days,they can attach to 80% of the bottom.Medicated serum with straw yellow and without deposition may be obtained by simple and mature methods. The results showed that the five kind of Bushen herhbs medicated serum all promoted the prolification of BMSCs.Osteogenesis inducer was positive control group,by ALP assay after culturing 2 generations,It reported that five kinds of Bushen herbs didn't induce the differentiation towards osteoblasts of marrow strowal cell of BMSCs.ConclusionsPrepare the drug-containing serum of five kind of Bushen herbs such as Herba Epimedii,morinda officinalis How,Eucommia ulmoides Oliv,Fructus Psoraleae and Herba Cistanches respectively and cultured the BMSCs with them. The results showed that all of them promoted the prolification of the BMSCs in different degree while on differentiation,there were no positive results. The main mechanism of clinic treating on racture,osteoporesis and osteoarthritis with tranditional Chinese medicine are the promotion of Bushen herbs on BMSCs. |
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