Axonal growth of embryonic stem cell-derived motoneurons in vitro and in motoneuron-injured adult rats
Axonal growth of embryonic stem cell-derived motoneurons in vitro and in motoneuron-injured adult rats
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DOI：
10.1073/pnas.0401103101
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发表时间：
2004-05-04
影响因子：
11.1
通讯作者：
Kerr, DA
中科院分区：
文献类型：
--
作者：
Harper, JM;Krishnan, C;Kerr, DA
我们从胚胎干细胞（ES）中产生脊髓运动神经元，以确定这些细胞在体外的发育潜力和它们取代成年哺乳动物脊髓运动神经元的能力。ES细胞来源的运动神经元延长长轴突，形成神经肌肉接头，并诱导肌肉收缩时，与成肌细胞共培养。我们将运动神经元定向的ES细胞移植到运动神经元损伤的成年大鼠的脊髓中，发现大约3,000个ES细胞衍生的运动神经元（输入量的25%）在每只动物的脊髓中存活>1个月。ES细胞衍生的轴突生长被髓磷脂抑制，并且这种抑制通过在体外和体内施用二丁酰cAMP（dbcAMP）或Rho激酶抑制剂来克服。在输注dbcAMP的移植大鼠中，在每只动物的前根内观察到约80个ES细胞来源的运动轴突，而在未用dbcAMP处理的移植大鼠中没有观察到。由于这些细胞复制了许多真正运动神经元的发育和成熟特征，因此它们是了解体外运动单位形成的重要生物学工具，也是重建体内神经回路的潜在治疗工具。
We generated spinal motoneurons from embryonic stem (ES) cells to determine the developmental potential of these cells in vitro and their capacity to replace motoneurons in the adult mammalian spinal cord. ES cell-derived motoneurons extended long axons, formed neuromuscular junctions, and induced muscle contraction when cocultured with myoblasts. We transplanted motoneuron-committed ES cells into the spinal cords of adult rats with motoneuron injury and found that approximate to3,000 ES cell-derived motoneurons (25% of input) survived for >1 month in the spinal cord of each animal. ES cell-derived axonal growth was inhibited by myelin, and this inhibition was overcome by administration of dibutyryl cAMP (dbcAMP) or a Rho kinase inhibitor in vitro and in vivo. In transplanted rats infused with dbcAMP, approximate to80 ES cell-derived motor axons were observed within the ventral roots of each animal, whereas none were observed in transplanted rats not treated with dbcAMP. Because these cells replicate many of the developmental and mature features of true motoneurons, they are an important biological tool to understand formation of motor units in vitro and a potential therapeutic tool to reconstitute neural circuits in vivo.