Name: a co-culture method to study neurite outgrowth in response to dental pulp paracrine signals
Uploaded: 2020-02-14T14:00:00
Description: Watch this Scientific Journal Video about A Co-Culture Method to Study Neurite Outgrowth in Response to Dental Pulp Paracrine Signals at JoVE.com

这项工作得到了国家卫生研究院/NIAMS（授予编号R01 AR062507和R01 AR053860至RS）的支持，b） 阿拉巴马大学伯明翰牙科学术研究培训（DART）赠款（编号T90DE022736（PI MacDougall）从国家牙科和颅面研究所/国家卫生研究院向SBP提供， c） UAB全球颅面、口腔和牙科疾病中心（GC-CODE）试点和可行性赠款给SBP和d）国家牙科和颅面研究所研究/国家卫生研究院K99 DE024406赠款给SBP。

所有小鼠实验均获得UAB机构动物护理和使用委员会（IACUC）的批准。
1. 板材制备
注： 盖玻片可用于在测定结束时对 DP 细胞进行成像。在无菌组织培养罩外的所有孵育和冲皮步骤期间，确保板盖已打开，以防止样品处理过程中受到污染。
<ol><li>
盖玻片准备
	<ol>
<li>高压灭菌器圆形盖玻片。</li>
		<li>过滤引擎盖下的超纯水。</li>
		<li>将盖玻片?...

<ol>
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W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimitotic+drugs+that+enhance+neuronal+survival+in+olfactory+bulb+cell+cultures.">Antimitotic drugs that enhance neuronal survival in olfactory bulb cell cultures.</a> Brain Research. 261 (2), 261-275 (1983).</li><li>Katzenell, S., Cabrera, J. R., North, B. J., Leib, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation,+Purification,+and+Culture+of+Primary+Murine+Sensory+Neurons.">Isolation, Purification, and Culture of Primary Murine Sensory Neurons.</a> Methods in molecular biology. 1656, 229-251 (2017).</li><li>Dussor, G. O., Price, T. J., Flores, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activating+transcription+factor+3+mRNA+is+upregulated+in+primary+cultures+of+trigeminal+ganglion+neurons.">Activating transcription factor 3 mRNA is upregulated in primary cultures of trigeminal ganglion neurons.</a> Molecular Brain Research. 118 (1-2), 156-159 (2003).</li><li>Lillesaar, C., Arenas, E., Hildebrand, C., Fried, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+rat+trigeminal+neurones+to+dental+pulp+cells+or+fibroblasts+overexpressing+neurotrophic+factors+in+vitro.">Responses of rat trigeminal neurones to dental pulp cells or fibroblasts overexpressing neurotrophic factors in vitro.</a> Neuroscience. 119 (2), 443-451 (2003).</li><li>Lillesaar, C., Eriksson, C., Fried, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+tooth+pulp+cells+elicit+neurite+growth+from+trigeminal+neurones+and+express+mRNAs+for+neurotrophic+factors+in+vitro.">Rat tooth pulp cells elicit neurite growth from trigeminal neurones and express mRNAs for neurotrophic factors in vitro.</a> Neuroscience Letters. 308 (3), (2001).</li><li>Lillesaar, C., Eriksson, C., Johansson, C. S., Fried, K., Hildebrand, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tooth+pulp+tissue+promotes+neurite+outgrowth+from+rat+trigeminal+ganglia+in+vitro.">Tooth pulp tissue promotes neurite outgrowth from rat trigeminal ganglia in vitro.</a> Journal of neurocytology. 28 (8), 663-670 (1999).</li><li>Chmilewsky, F., Ayaz, W., Appiah, J., About, I., Chung, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nerve+Growth+Factor+Secretion+From+Pulp+Fibroblasts+is+Modulated+by+Complement+C5a+Receptor+and+Implied+in+Neurite+Outgrowth.">Nerve Growth Factor Secretion From Pulp Fibroblasts is Modulated by Complement C5a Receptor and Implied in Neurite Outgrowth.</a> Scientific reports. 6, 31799 (2016).</li><li>Pagella, P., Neto, E., Jim&#195;&#169;nez-Rojo, L., Lamghari, M., Mitsiadis, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidics+co-culture+systems+for+studying+tooth+innervation.">Microfluidics co-culture systems for studying tooth innervation.</a> Frontiers in Physiology. 5, 326 (2014).</li><li>Miura, T., Yokokawa, R. <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+on+a+chip:+Developmental+biology+applications+of+self-organized+capillary+networks+in+microfluidic+devices.">Tissue culture on a chip: Developmental biology applications of self-organized capillary networks in microfluidic devices.</a> Development, Growth &amp; Differentiation. 58 (6), 505-515 (2016).</li></ol>

口腔的日常活动要求牙齿感知外部刺激和内部炎症，以便正确使用和维护。然而，关于驱动牙齿内侧发育过程的信号，只有有限的信息可用。该协议提供了一种分离和共同培养原DP细胞和TG神经元的方法，以研究两个群体之间的交叉通信。如下所述，对几个变量进行了优化，并留下了进一步的研究渠道。
控制在本次测定的每一步都很重要。每个测定中都应包含具有无基础DP细?...

牙齿内侧使牙齿能够感知压力、温度和炎症，所有这些都对牙齿器官的使用和维护至关重要。未能感觉到与蛀牙和创伤相关的牙痛会导致疾病进展。因此，适当的内侧是正常牙齿生长、功能和护理的必要要求。
虽然大多数器官在出生时功能齐全，内在，牙齿发育延伸到成年，牙齿内侧和矿化在产后阶段1、2中协同发生。有趣的是，牙髓（DP）间质在胚胎形成过程中最初分泌排斥信号，以防止斧头进入发育中的牙齿器官，后来随着牙齿接近爆发3、4，它转移到吸引因子的分泌。在产后阶段，三角 （TG） 神经的一个发泡的斧头在牙本质沉积开始的时间渗透到牙齿中并贯穿整个牙齿（在 Pagella，P. 等人 5中进行了审查）。几项体内研究表明，神经元间质相互作用指导小鼠的牙齿内侧（在Luukko，K.等人6中回顾），但分子机制的细节很少。
细胞共培养物提供受控环境，调查人员可以在其中操纵神经元和间质体之间的相互作用。共培养实验使得深入探讨引导牙齿内向和发展的信号通路成为可能。然而，用于研究共培养细胞的几种常规方法带来了技术挑战。例如，神经酸盐生长的水晶紫罗兰染色可以非特有染色施万细胞包含在TG捆绑分散体中，并且可能存在颜色强度的峰值，反应相对较小7。微流体室提供了一个有吸引力的选择，但比跨井过滤器8，9昂贵得多，并且只允许研究神经元对DP分泌物的反应。为了解决这些问题，我们开发了一个协议，允许：a） 对TG神经质生长的精确染色和成像，以响应DP分泌物;b） 对DP细胞和/或TG神经元进行基因改造，以研究特定的信号通路，以及c）研究DP细胞对TG神经元分泌因子的反应。该协议提供了在体外共培养测定的受控环境中精确研究牙齿内侧的几个特征的能力。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>5-Fluoro-2&#39;-deoxyuridine</td>
			<td>Sigma-Aldrich</td>
			<td>F0503</td>
			<td>Used as a mitotic Inhibitor at 15 &#956;M concentration in co-culture media, Day 2</td>
		</tr>
		<tr>
			<td>24 Well Cell Culture Plate</td>
			<td>Corning</td>
			<td>3524</td>
			<td>Co-culture plate</td>
		</tr>
		<tr>
			<td>Alexa-546 anti-chicken</td>
			<td>Invitrogen</td>
			<td>A-11040</td>
			<td>Secondary to stain neurite outgrowth labeled by anti-GFP antibody, 1:500 dilution</td>
		</tr>
		<tr>
			<td>Anti-GFP Antibody</td>
			<td>Aves Lab, Inc</td>
			<td>GFP-1010</td>
			<td>Primary antibody to label Thy1-YFP neurons, 1:200 dilution</td>
		</tr>
		<tr>
			<td>Anti-Neurofilament 200 antibody</td>
			<td>Sigma-Aldrich</td>
			<td>NO142</td>
			<td>Monoclonal primary antibody to label neurons, 1:1000 dilution, alternative if YFP mice are not available</td>
		</tr>
		<tr>
			<td>B6;129- Tgfbr2tm1Karl/J</td>
			<td>The Jackson Laboratory</td>
			<td>12603</td>
			<td>Tgfbr2f/f mouse model used for dental pulp cells in optimized protocol</td>
		</tr>
		<tr>
			<td>B6.Cg-Tg(Thy1-YFP)16Jrs/J</td>
			<td>The Jackson Laboratory</td>
			<td>3709</td>
			<td>Thy1-YFP mouse model genotype used for trigeminal neurons</td>
		</tr>
		<tr>
			<td>Collagenase Type II</td>
			<td>Millipore</td>
			<td>234155-100MG</td>
			<td>Used to disperse trigeminal neurons</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Gibco</td>
			<td>10437</td>
			<td>Additive to co-culture media</td>
		</tr>
		<tr>
			<td>Fine forceps</td>
			<td>Fine Science Tools</td>
			<td>11413-11</td>
			<td>Fine forceps for TG dissection</td>
		</tr>
		<tr>
			<td>Laminin</td>
			<td>Sigma-Aldrich</td>
			<td>L2020</td>
			<td>Coats the transwell inserts at final concentration of 10 &#956;g/ml, stock solution is assumed at 1.5 mg/ml</td>
		</tr>
		<tr>
			<td>Lysis Buffer (Buffer RLT)</td>
			<td>Qiagen</td>
			<td>79216</td>
			<td>Extracts RNA from dental pulp cells post co-culture</td>
		</tr>
		<tr>
			<td>L-Glutamine</td>
			<td>Gibco</td>
			<td>25030081</td>
			<td>Additive to co-culture media</td>
		</tr>
		<tr>
			<td>Micro-dissecting scissors</td>
			<td>Sigma-Aldrich</td>
			<td>S3146-1EA</td>
			<td>Dissection scissors to open skull</td>
		</tr>
		<tr>
			<td>Microscope Cover Glass</td>
			<td>Fisherbrand</td>
			<td>12-545-81</td>
			<td>Circlular coverslip for optional cell culturing and immunofluorescence processing</td>
		</tr>
		<tr>
			<td>Minimal Essential Medium a</td>
			<td>Gibco</td>
			<td>12571063</td>
			<td>Co-culture media base</td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin</td>
			<td>Gibco</td>
			<td>15070063</td>
			<td>Antibiotic additive to co-culture media</td>
		</tr>
		<tr>
			<td>Phosphatase Inhibitor</td>
			<td>Sigma-Aldrich</td>
			<td>04 906 837 001</td>
			<td>Additive to RIPA Buffer for extracting protein from dental pulp cells post co-culture</td>
		</tr>
		<tr>
			<td>Polybrene</td>
			<td>Millipore</td>
			<td>TR-1003-G</td>
			<td>Used to aid in dental pulp cell transfection</td>
		</tr>
		<tr>
			<td>Poly-D-Lysine</td>
			<td>Sigma-Aldrich</td>
			<td>P7280</td>
			<td>Coverslip coating to aid dental pulp cellular adhesion</td>
		</tr>
		<tr>
			<td>Protease Inhibitors</td>
			<td>Millipore</td>
			<td>05 892 791 001</td>
			<td>Additive to RIPA Buffer for extracting protein from dental pulp cells post co-culture</td>
		</tr>
		<tr>
			<td>RNAse/DNAse free eppendorf tubes</td>
			<td>Denville</td>
			<td>C-2172</td>
			<td>Presterilized 1.7 ml tubes for RNA, DNA or protein collection at the end of assay</td>
		</tr>
		<tr>
			<td>ThinCert Cell Culture Insert</td>
			<td>Greiner Bio-One</td>
			<td>662631</td>
			<td>Transwell inserts for trigeminal neurons in co-culture assays</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.25%)</td>
			<td>Gibco</td>
			<td>25200056</td>
			<td>Used fto disperse dental pulp cells</td>
		</tr>
		<tr>
			<td>Trypsin Type II</td>
			<td>Sigma-Aldrich</td>
			<td>T-7409</td>
			<td>Used to disperse trigeminal neurons</td>
		</tr>
		<tr>
			<td>Ultra Fine Forceps</td>
			<td>Fine Science Tools</td>
			<td>11370-40</td>
			<td>Ultra fine forceps for dissection</td>
		</tr>
		<tr>
			<td>Uridine</td>
			<td>Sigma-Aldrich</td>
			<td>U3750</td>
			<td>Used as a mitotic Inhibitor at 1 &#956;M concentration in co-culture media, Day 2</td>
		</tr>
		<tr>
			<td>Vacuum Filtration System</td>
			<td>Millipore</td>
			<td>SCNY00060</td>
			<td>Steriflip disposable filter, 50 &#956;m nylon net filter</td>
		</tr>
		<tr>
			<td>Vial forceps</td>
			<td>Fine Science Tools</td>
			<td>110006-15</td>
			<td>Long forceps for tissue transfer to conicals</td>
		</tr>
	</tbody>
</table>

a co-culture method to study neurite outgrowth in response to dental pulp paracrine signals

牙齿内侧使牙齿能够感知压力、温度和炎症，所有这些都对牙齿器官的使用和维护至关重要。没有感官内向，日常的口腔活动将造成无法弥补的损害。尽管它很重要，但内在牙齿在牙齿发育和维护中的作用在很大程度上被忽视了。几项研究表明，DP细胞分泌细胞外基质蛋白和副体信号，以吸引和引导TG斧头进入牙齿并在整个牙齿中。然而，很少有研究对DP间质和神经元亲和力之间的相声有详细的见解。为了解决知识的这一差距，研究人员已经开始利用共同文化和各种技术来研究这些相互作用。在这里，我们演示了在具有大直径孔隙的覆盖跨孔过滤器上分散的TG神经元共同培养初级DP细胞的多个步骤，以允许斧孔通过孔隙生长。利用腺病毒-Cre-GFP重组酶系统，利用具有致利益基因的初级DP细胞，利用loxP位点促进基因删除。使用 Thy1-YFP 鼠标中的 TG 神经元可以进行精确的对焦成像，通过共聚焦显微镜的表达远高于背景水平。DP反应可以通过蛋白质或RNA的收集和分析，或者通过免疫荧光染色的DP细胞镀在可移动玻璃盖玻片。介质可以使用技术，如蛋白组分析进行分析，虽然这将需要白蛋白耗尽，因为胎儿牛血清在介质中的存在。该协议提供了一个简单的方法，可以操纵研究TG神经元和DP细胞的形态、遗传和细胞骨骼反应，以响应共培养测定的受控环境。

这些结果表明，与TG神经酸盐单一培养的控制相比，TG神经酸盐出生长在基础井中原DP细胞的存在下增加（图2A，C）。在神经质外生长中有一些测定到测定的变异性。因此，TG神经元单一培养应包含在所有测定中，作为检测神经质外生长基底水平的控制。在Ad-Cre-GFP和Ad-eGFP感染后，Tgfbr2f/f小鼠的初级细胞被用于此协议（图2D）。<...

Watch this Scientific Journal Video about A Co-Culture Method to Study Neurite Outgrowth in Response to Dental Pulp Paracrine Signals at JoVE.com

A Co-Culture Method to Study Neurite Outgrowth in Response to Dental Pulp Paracrine Signals

我们描述了牙髓（DP）原细胞的分离、分散和电镀，这些细胞在覆盖的透井过滤器上培养三叉（TG）神经元。DP细胞的细胞反应可以通过免疫荧光或RNA/蛋白质分析进行分析。神经元标记物与共聚焦显微镜的免疫荧光允许分析神经质外生长反应。

研究对牙髓寄生虫信号反应的神经质生长的共同培养方法

Tooth innervation allows teeth to sense pressure, temperature and inflammation, all of which are crucial to the use and maintenance of the tooth organ. ...

该协议提供了如何与三叉神经元共同培养牙髓干细胞的详细大纲。有了它，我们可以调查由他们的相声驱动的多个响应。这种技术的主要优点是能够研究和操作细胞群体之一或两个细胞群，并精确测量结果。这种方法与神经研究有直接关系，可以深入了解牙髓干细胞如何修复创伤性事件或神经退行性疾病受损的神经组织。这种技术有几个阶段要学习，很难把它们全部保留一些。该协议详细介绍了每个阶段，以帮助这一点。牙髓和牙髓难以分散，需要各种涡流和移液。即使这样，您也无法获得完全分散，因此需要优化和复制。对鼠标实施安乐死后，将头部放在一次性垫上，使嘴朝向天花板，颈部底座平放在工作表面上。使用剃须刀刀片和锯切运动将毛细剑与 maxilla 分开。用剪刀或钳子去除舌头，以便更容易地接触摩尔。将开放的头放在无菌纱布垫上的盘子中，将试样放在解剖显微镜下。取出第一个摩尔周围的肺骨组织。将钳子插入肺泡开口，将组织从牙齿转向嘴上的水桶或语言侧。收集所有最大第一摩尔，并轻轻地将亚曼地和最大摩尔转移到一个单独的细胞培养皿与1X PBS在冰上。去除每个大摩尔外侧周围的釉质外器官。使用一组钳子旋转摩尔，使尖子向下，打开的根被公开。牙齿底部有一个椭圆形开口，不透明的牙髓组织被薄薄的牙龈和牙釉质层封装。使用钳子的尖端，通过围绕矿化组织的内部周长运行一臂钳，轻轻松开牙髓。从矿化结构中去除牙髓组织，转移到含有 1X PBS 的第三个培养皿中。如果釉质外器官尚未分离，请将其取出。在 50 毫升锥形管中将所有牙髓组织转移到 0.25% 的 trypsin EDTA 中。漩涡混合物，并放在一个37摄氏度的温水浴10分钟。在无菌罩下，将加热的共培养基介质添加到至少一对一介质的最终比例中，以尝试性酶来灭活酶。用10毫升移液器多次上下移料，以进一步分散介质中的牙髓，避免出现大气泡。将分散的牙髓的一毫升转移到24井组织培养板的每个井中。将板放在摄氏37度的培养箱中。并允许细胞在改变介质之前，从未分散的组织上连接并迁移48小时。安乐死和去除皮肤后，将一把微型解剖剪刀的尖端插入头骨底部。沿着头骨的缝合线切割。使四个小的水平切口沿冠状缝合线的耳朵，直到沿着头骨底部的羔羊缝合线。这将创建两个骨瓣。用钳子剥回两个骨瓣，露出大脑。找到位于大脑和骨骼之间的三叉神经。切开三根树枝，这些树枝会向眼睛、最大和毛细血管中游去。并使用直边细钳将刚将钢联转移到冰盘上的冷 1X PBS。一旦收获所有三叉束使用文件钳将刚龙转移到50毫升锥形管，每毫升含有5毫克的无菌过滤胶原酶II.Vortex混合物，将管放在37摄氏度的水浴中25至30分钟。每五到十分钟，把管子从水浴中拿出来，漩涡，然后回到浴池里。在643xg下将胶原酶三叉神经元溶液离心两分钟。在组织培养罩下，用微管轻轻吸吸胶原酶，加入5毫升1%无菌过滤的三普辛II型，然后将管子放入37摄氏度的水浴中25至30分钟，并在这段时间内每隔5分钟短暂旋转一次管。以 trypsin 与介质的一对一比例添加媒体，以停用剩余的 trypsin。计算细胞数量，在介质中稀释混合物至每毫升20万个细胞。将涂层的转井过滤器放入带牙髓的井中。将250微升的移液器放到转井过滤器上，并在37摄氏度的温度下培养细胞。第二天，用一毫升共培养基介质代替介质，辅以一微摩尔尿素和15微摩尔5-氟-2'脱氧核糖核酸，以阻止可能防止神经性细胞生长的间质细胞过度增殖。你必须添加线粒体抑制剂的第二天或神经素生长不会发生。在这项研究中，与控制三叉神经性单培养相比，在底层井中存在原生牙髓细胞时，三叉神经土的生长增加。从TGF-beta受体2浮子/flox小鼠分离的原细胞在注射ade-Cre-GFP或ade-eGFP后数量相等。半定量PCR表明，腺病毒-Cre-GFP删除了侧翼基因转化生长因子β受体2，腺病毒eGFP作为控制病毒载体。在生长因子转化β受体2缺失的培养中，神经素的外生长量减少。细胞群在晶体紫罗兰染色后对透井过滤器进行亮场成像，表明大孔隙普遍存在。大箭头指向具有中质形态的细胞，而小箭头指向神经元形态的细胞。水晶紫罗兰没有偏置地染色了两个细胞。此外，使用alex488二级抗体的β-III管状细胞的免疫荧光染色显示多个细胞的非特异性染色，使得对外向结构的成像变得困难。因为牙髓细胞和三叉神经元都不易分散，所以每个研究人员都需要优化他们的电镀程序。如果您单独在单独的井中培养牙髓细胞，您可以使用免疫荧光，或者您可以量化RNA或蛋白质水平，以确定牙髓细胞对神经元的反应。记得漂白任何与腺病毒接触的容器或提示。一定要把剃须刀正确丢弃在锋利的垃圾箱里。

Research

JoVE Journal

Developmental Biology

An Optogenetic Approach for Assessing Formation of Neuronal Connections in a Co-culture System

评定神经连接的形成在共培养体系的光遗传学方法

Here we describe a protocol to generate a co-culture consisting of 2 different neuronal populations. Induced pluripotent stem cells (iPSCs) are ...

科研

发育生物学

Generating CRISPR/Cas9 Mediated Monoallelic Deletions to Study Enhancer Function in Mouse Embryonic Stem Cells

生成CRISPR / Cas9介导的单等位基因缺失来研究小鼠胚胎干细胞功能增强

Enhancers control cell identity by regulating tissue-specific gene expression in a position and orientation independent manner. These enhancers are often ...

Using Touch-evoked Response and Locomotion Assays to Assess Muscle Performance and Function in Zebrafish

采用触摸诱发反应和运动测定以评估在斑马鱼肌肉性能和功能

Zebrafish muscle development is highly conserved with mammalian systems making them an excellent model to study muscle function and disease. Many ...

Application of Impermeable Barriers Combined with Candidate Factor Soaked Beads to Study Inductive Signals in the Chick

在小鸡学习感应信号与候选因子浸泡珠结合防渗屏障中的应用

The chick embryo provides a superb vertebrate model that can be used to dissect developmental questions in a direct way. Its accessibility and robustness ...

The Aortic Ring Co-culture Assay: A Convenient Tool to Assess the Angiogenic Potential of Mesenchymal Stromal Cells In Vitro

The Aortic Ring Co-culture Assay: A Convenient Tool to Assess the Angiogenic Potential of Mesenchymal Stromal Cells In Vitro

主动脉环共培养法: 一种方便的工具, 可以评估骨髓间质细胞的血管生成潜能体外

Angiogenesis is a complex, highly regulated process responsible for providing and maintaining adequate tissue perfusion. Insufficient vasculature ...

Organotypic Culture Method to Study the Development Of Embryonic Chicken Tissues

脊髓培养法研究胚胎鸡组织的发展

The embryonic chicken is commonly used as a reliable model organism for vertebrate development. Its accessibility and short incubation period makes it ...

An In Vivo Method to Study Mouse Blood-Testis Barrier Integrity

An In Vivo Method to Study Mouse Blood-Testis Barrier Integrity

小鼠血睾丸屏障完整性的一种活体研究方法

Spermatogenesis is the development of spermatogonia into mature spermatozoa in the seminiferous tubules of the testis. This process is supported by ...

Isolation, Propagation, and Prion Protein Expression During Neuronal Differentiation of Human Dental Pulp Stem Cells

人牙髓干细胞神经分化过程中的分离、繁殖及普利翁蛋白表达

Bioethical issues related to the manipulation of embryonic stem cells have hindered advances in the field of medical research. For this reason, it is very ...

Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C (UV-C) Damage

Assessment of Oxidative Damage in the Primary Mouse Ocular Surface Cells/Stem Cells in Response to Ultraviolet-C UV-C Damage

评估原小鼠眼部表面细胞/干细胞对紫外线-C （UV-C） 损伤的反应

The ocular surface is subjected to regular wear and tear due to various environmental factors. Exposure to UV-C radiation constitutes an occupational ...

An Enhanced Green Fluorescence Protein-based Assay for Studying Neurite Outgrowth in Primary Neurons

增强的绿色荧光蛋白测定，用于研究原神经元的神经质生长

Neurite outgrowth is a fundamental event in the formation of the neural circuits during nervous system development. Severe neurite damage and synaptic ...