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Differentiation of Oligodendrocytes from Mouse Induced Pluripotent Stem Cells Without Serum

机译：从小鼠诱导的无血清多能干细胞中分化少突胶质细胞

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摘要

Cell therapy using induced pluripotent stem (iPS) cells might become a new approach for treating neonatal hypoxic–ischemic injury such as periventricular leukomalacia. To obtain appropriate donor cells for transplantation, we differentiated oligodendrocyte (OL) lineage cells from mouse iPS cells. Induction of OL lineage cell differentiation from iPS cells was carried out with a seven-step culture method. Mouse iPS cells (stage 1) were induced to form embryoid bodies for 4 days under a serum-free condition that was suitable for ectoderm induction (stage 2), following by selection of nestin-positive neural stem cells (NSCs) for 10–12 days (stage 3). NSCs were cultured in expansion medium containing fibroblast growth factor (FGF)-2 for 4 days (stage 4), induced to differentiate into glial progenitor cells by epidermal growth factor and fibroblast growth factor (FGF-2) treatment for 4–5 days (stage 5), and then into OL progenitor cells by culture in neurobasal A medium containing FGF-2 and platelet-derived growth factor for 6–8 days (stage 6). Terminal differentiation into O4-positive OLs was carried out by culture in neurobasal A containing T3 and ciliary neurotrophic factor for 7 days (stage 7). Inwardly rectifying K+ currents, which are characteristic of OLs, were detected in iPS cell-derived cells at stage 7 in whole cell clamp mode. Our data suggest that OLs can be effectively differentiated from mouse iPS cells without serum in a stepwise manner, which may be appropriate for use as donor cells in transplantation.
机译：使用诱导性多能干（iPS）细胞进行细胞疗法可能成为治疗新生儿缺氧缺血性损伤（如脑室白细胞减少症）的新方法。为了获得合适的供体细胞进行移植，我们从小鼠iPS细胞中分化出少突胶质细胞（OL）谱系细胞。用七步培养法从iPS细胞诱导OL谱系细胞分化。在适合于外胚层诱导的无血清条件下，将小鼠iPS细胞（第1期）诱导形成胚状体4天（第2期），然后选择巢蛋白阳性神经干细胞（NSC）10-12天（第3阶段）。 NSC在含有成纤维细胞生长因子（FGF）-2的扩增培养基中培养4天（第4阶段），通过表皮生长因子和成纤维细胞生长因子（FGF-2）处理诱导分化为神经胶质祖细胞4-5天（阶段5），然后在含有FGF-2和血小板衍生生长因子的神经基底A培养基中培养6-8天，进入OL祖细胞（阶段6）。通过在含有T3和睫状神经营养因子的神经基底A中培养7天来最终分化为O4阳性OL（第7阶段）。在全细胞钳制模式下，在第7阶段在iPS细胞衍生的细胞中检测到了向内整流的K +电流，这是OL的特征。我们的数据表明，可以在没有血清的情况下以逐步方式将OLs与小鼠iPS细胞有效区分，这可能适合用作移植中的供体细胞。

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  • 来源
    《Translational Stroke Research》 |2013年第2期|149-157|共9页
  • 作者

    Sachiyo Misumi; Ruriko Nishigaki; Yoshitomo Ueda; Yoko Watanabe; Yuko Shimizu; Akimasa Ishida; ChaGyun Jung; Hideki Hida;

  • 作者单位

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

    Department of Neurophysiology and Brain Science Nagoya City University Graduate School of Medical Sciences">(1);

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  • 正文语种 eng
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  • 关键词

    Periventricular leukomalacia; Donor cells; Embryoid body; Inwardly rectifying current; O4 antigen;

    机译：脑室周围白血球减少症;供体细胞;胚状体;内向整流电流;O4抗原;

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