Name: study glial cell heterogeneity influence on axon growth using a new coculture method
Uploaded: 2010-09-06T17:00:00
Description: Watch this Scientific Journal Video about Study Glial Cell Heterogeneity Influence on Axon Growth Using a New Coculture Method at JoVE.com

这项研究是FMMU新发现的基础和部分美国国立卫生研究院拨款的支持。

1。胶质细胞培养来自不同地区的中枢神经系统神经胶质细胞可以培养。整个过程是在过程图所示。 <img src="/files/ftp_upload/2111/2111process.jpg" alt="过程1" />第1天涂层培养板和盖玻片<ol><li>干燥灭菌的玻璃显微镜下一轮盖玻片高压灭菌器。 </li><li>板到灭菌的24孔培养板消毒盖玻片。 </li><li>外套与多聚赖氨酸和根的温度下2小时孵化的盖玻片。...

<ol>
	<li>Rossignol, S., Schwab, M., Schwartz, M., Fehlings, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+injury:+time+to+move?.">Spinal cord injury: time to move?.</a> J Neurosci. 27 (44), 11782-11792 (2007).</li><li>Yiu, G., He, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glial+inhibition+of+CNS+axon+regeneration.">Glial inhibition of CNS axon regeneration.</a> Nat Rev Neurosci. 7 (8), 617-627 (2006).</li><li>Barres, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mystery+and+magic+of+glia:+a+perspective+on+their+roles+in+health+and+disease.">The mystery and magic of glia: a perspective on their roles in health and disease.</a> Neuron. 60 (3), 430-440 (2008).</li><li>Matyash, V., Kettenmann, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heterogeneity+in+astrocyte+morphology+and+physiology.">Heterogeneity in astrocyte morphology and physiology.</a> Brain Res Rev. , (2009).</li><li>Wanner, I. B., Deik, A., Torres, M., Rosendahl, A., Neary, J. T., Lemmon, V. P., Bixby, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+in+vitro+model+of+the+glial+scar+inhibits+axon+growth.+Glia.">A new in vitro model of the glial scar inhibits axon growth. Glia.</a> 56 (15), 1691-1709 (2008).</li><li>Kuffler, D. P., Sosa, I. J., Reyes, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Schwann+cell+chondroitin+sulfate+proteoglycan+inhibits+dorsal+root+ganglion+neuron+neurite+outgrowth+and+substrate+specificity+via+a+soma+and+not+a+growth+cone+mechanism.">Schwann cell chondroitin sulfate proteoglycan inhibits dorsal root ganglion neuron neurite outgrowth and substrate specificity via a soma and not a growth cone mechanism.</a> J Neurosci Res. 87 (13), 2863-2871 (2009).</li><li>Kleitman, N., Wood, P. M., Bunge, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+culture+method+for+the+study+of+myelination.">Tissue culture method for the study of myelination.</a> Culturing nerve cells. , P559-P559 (1998).</li></ol>

这个实验协议的目的是要达到两个目标，以研究神经胶质细胞，神经细胞粘附和突起的生长，特别是星形胶质细胞的异质性的影响。第一个目标是保持星形胶质细胞的异质性，尽可能在本实验中，胶质细胞的融合文化，丰富的星形胶质细胞混合小学文化，无任何化学处理和消化的传播，这可能会进一步破坏细胞，并诱导星形胶质细胞的损伤反应。最后的星形胶质细胞纯度为90％以上，GFAP阳性污染?...

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		<th>Catalogue Number</th>
		<th>Comment</th>
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<td>DMEM(high glucose)</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>10313-039</td>
<td>&nbsp;</td>
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<td>L15 medium</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>11415-114</td>
<td>&nbsp;</td>
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<td>FBS</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>10437-077</td>
<td>&nbsp;</td>
<tr>
<td>Neurobasal Medium</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>21103-049</td>
<td>&nbsp;</td>
<tr>
<td>poly-lysine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>P4832</td>
<td>culture grade</td>
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<td>NGF(2.5S)</td>
<td>&nbsp;</td>
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<td>13257-019</td>
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<td>17504-044</td>
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<td>Invitrogen</td>
<td>25200-056</td>
<td>&nbsp;</td>
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<td>FUDR</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>F0503</td>
<td>&nbsp;</td>
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<td>neurofilament antibody</td>
<td>&nbsp;</td>
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<td>ab24575</td>
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study glial cell heterogeneity influence on axon growth using a new coculture method

在中央的所有哺乳动物的中枢神经系统，切断损伤后的轴突无法再生到原来的目标和功能恢复是非常差的1。轴突再生的失败是几个因素，包括敌对的神经胶质细胞环境，抑制髓鞘相关分子和减少内在的神经元的再生能力2的综合作用的结果。星形胶质细胞是最突出的中枢神经系统的神经胶质细胞类型，并在轴突功能的生理和病理条件3下发挥重要作用。对比同源少突胶质细胞，星形胶质细胞是一个由不同的星形胶质细胞亚群具有不同的形态和基因表达的 4组成的异质细胞群。这种异质性的功能意义，如对轴突的生长影响，在很大程度上是未知的。 为了研究神经胶质细胞，尤其是在星形胶质细胞神经​​元行为的异质性的功能，我们建立了合作培养高纯度的背根与从大鼠皮层获得的神经胶质细胞神经​​节神经元的一种新方法。通过这项技术，我们能够直接比较不同的星形胶质细胞亚群在相同条件下神经细胞的粘附和轴突的生长。 在这份报告中，我们给这种方法的星形胶质细胞的分离和文化，背根神经节神经元的分离和提纯，和背根神经节神经元与星形胶质细胞共培养的详细协议。这种方法也可以扩展到其他脑区，研究细胞或地区的特定神经元和神经胶质细胞之间的相互作用。

Watch this Scientific Journal Video about Study Glial Cell Heterogeneity Influence on Axon Growth Using a New Coculture Method at JoVE.com

Study Glial Cell Heterogeneity Influence on Axon Growth Using a New Coculture Method

在这个协议中，我们描述了一个新的方法来研究与轴突生长的神经胶质细胞的异质性的影响<em&gt;在体外</em&gt;共培养体系。大鼠皮层神经胶质细胞培养汇合和高度纯化的大鼠背根神经节神经元共培养。在相同的文化，不同的神经胶质细胞对神经细胞的粘附和轴突的生长影响比较直接。这种方法提供了一种新的方式直接研究胶质细胞对神经细胞粘附和轴突生长的异质性影响。

研究胶质细胞轴突的生长异质性的影响，使用一个新的共培养法

In the central nervous system of all mammals, severed axons after injury are unable to regenerate to their original targets and functional recovery is ...

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