用于SD大鼠神经祖细胞增殖分化研究的体外培养法的建立

Establishment of sprague-dawley rat neural progenitor cells cultivation for proliferation and differentiation research

目的建立一种用于SD大鼠神经祖细胞(NPC)增殖分化研究的体外培养法,为深入研究其增殖分化奠定基础。方法机械法分离和传代新生SD大鼠海马组织NPC,应用特定培养基进行体外培养,BrdU标记后应用免疫荧光法观察鉴定NPC和分化后神经细胞。对机械法离解后的活细胞率差异进行比较。结果培养的NPCs可以在体外进行培养扩增,体外传代至第9代末仍可见Nestin抗体阳性的神经球形成,并且有分化为3种神经终末细胞的潜能。每次机械法分离后的NPC活细胞率的差异无统计学意义(P>0.05)。结论建立的SD大鼠NPC体外培养法更利于增殖分化的研究。

Objective To establish a cultivation on SD rat neural progenitor cells（NPCs） proliferation and differentiation for further study.Methods The NPCs were isolated from neonatal SD rat hippocampus and passaged by mechanical method.Then the NPCs cultured in special medium in vitro.After signed by BrdU,the NPCs and the differentiated neura1 terminal cells could be identified by the immunofluorescence.The rates of living cells were compared.Results The neonatal SD rats＇ NPCs could be cultured and proliferated in vitro and could show the Nestin-positive neurospheres at the end of passage 9.The cells had the ability to differentiate into three neural terminal cells.The differents between the rates of living NPCs were not significant（P0.05）.Conclusion The SD rats＇ NPCs cultivation is benefit to the research of NPC proliferation and differentiation.