| 摘要: |
| 目的:探讨成体嗅上皮神经干细胞(olfactory epithelium neural stem cells)的培养方法及其体外分化的多潜能性。方法:使用中性蛋白酶和Ⅰ型胶原酶分阶段酶解消化嗅上皮,富集嗅上皮神经干细胞,扩增细胞,用加入生长因子bFGF和EGF的无血清培养液促使其形成神经球。血清诱导神经球分化并用免疫细胞化学染色方法鉴定分化细胞。结果:原代培养的嗅上皮神经干细胞呈集落样增殖,集落中的细胞表达在体分子标记GBC2和细胞角蛋白5。传代细胞用含生长因子的无血清培养液培养可增殖形成神经球。用含血清的培养液可诱导神经球分化；从神经球迁移出的分化细胞具有很高的异质性,其中包含有TUJ1阳性的神经元、P75NGFR阳性的嗅鞘细胞、GFAP阳性的星形胶质细胞和NG2阳性的寡突胶前体细胞,同时也存在有nestin阳性的神经干细胞。结论:成体嗅上皮神经干细胞可体外培养增殖形成神经球,神经球细胞具有向神经元、星形胶质细胞和寡突胶前体细胞分化的潜能。 |
| 关键词: 嗅上皮 神经干细胞 细胞分化 多潜能性 |
| DOI：10.3724/SP.J.1008.2009.0985 |
| 投稿时间：2009-07-31修订日期：2009-08-31 |
| 基金项目:国家自然科学基金(30770657,30530240). |
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| Multipotency of cultured olfactory epithelium neural stem cells |
| GAO Liang,CAO Li,SU Zhi-da,ZHU Yan-ling,HE Cheng* |
| (Institute of Neuroscience,College of Basic Medical Sciences,Second Military Medical University,Shanghai 200433,China) |
| Abstract: |
| Objective:To investigate the culture method for adult olfactory epithelium neural stem cells and their multipotency during in vitro differentiation.Methods: Olfactory epithelium was sequentially digested with neutral protease and collagenase ⅠA,then the olfactory epithelium neural stem cells were enriched and amplified.Neurospheres were formed by culturing the cells with serum free medium containing bFGF and EGF.Differentiation of neurospheres was induced with medium containing serum; the differentiated cells were identified by immunocytochemistry.Results: Primary cultured olfactory epithelium neural stem cells proliferated and formed colonies.Cells in the colonies expressed the in vivo markers GBC2 and cytokeratin 5.Passaged cells proliferated and formed neurospheres when cultured with serum free medium containing bFGF and EGF.Medium containing serum induced the differentiation of neurospheres.TUJ1 positive neurons,P75NGFR positive olfactory ensheathing cells,GFAP positive astrocytes and NG2 positive oligodendrocyte progenitors were detected in the highly heterogeneous differentiated cells; besides,the cells also contained nestin positive neural stem cells.Conclusion: Cultured adult olfactory epithelium neural stem cells can proliferate and form neurospheres.The neurosphere cells have the multipotency to differentiate into neurons,astrocytes and oligodendrocyte progenitors. |
| Key words: olfactory epithelium neural stem cells cell differentiation multipotency |
