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体外分离大鼠骨髓间充质干细胞方法研究
METHODOLOGY STUDY ON ISOLATING BONE MARROW-DERIVED MESENCHYMAL STEM CELLS IN VITRO
【摘要】 目的确定分离骨髓间充质干细胞 (mesenchymalstemcells,MSCs)过程中的各种条件对获得MSCs群的影响。方法应用不同的分离液、离心温度、速度、培养液及不同品种胎牛血清等条件分离培养细胞并进行对比分析。结果使用 2种分离液 ,在相同离散密度时 ,得到的细胞群相同 ;相同离散密度的 2种分离液 ,不同离心温度、速度及时间 ,如果培养条件相同 ,获得的细胞群也相同 ;用含国产或进口胎牛血清的培养基进行培养所获细胞有明显的不同 ;改良的最小必需培养基 (Dulbecco′sminimumessentialmedium ,DMEM )中添加F12与不添加F12获得的细胞克隆群基本相同。结论无论使用那种分离液 ,只要其离散密度在 10 73g/L均能获得较纯的MSCs;在确定温度、时间、速度等条件下 ,确定基础培养基、选择合适的胎牛血清是取得实验成功的关键。
【Abstract】 ObjectiveTo test the effect of the conditions of isolating bone marrow-derived mesenchymal stem cells (MSCs) on the isolated cells. MethodsMSCs were obtained and cultured using different separation media,different centrifugal temperature,time,speed and different culture media, then the obtained data were analysed. ResultsWhen using two kinds of separation media of the same concentration MSCs isolated were the same, but different centrifugal temperature,time and speed, the isolated MSCs were also the same, while MSCs cultured in media containing domestic or imported fetal bovine serum(FBS) were obviously different. MSCs cultured in Dulbecco′s minimum essential medium(DMEM) added with F12 or not were almost the same.ConclusionWhichever separation medium in choice, relative pure MSCs are available if the concentration is at 1 073 g/L,can help obtain relatively pure MSCs. Under the same conditions of centrifugal temperature,time,speed,if the basic medium is defined it is critical to select optimal FBS.
- 【文献出处】 河北医科大学学报 ,Journal of Hebei Medical University , 编辑部邮箱 ,2004年03期
- 【分类号】R329.2
- 【被引频次】11
- 【下载频次】144