Name: in vitro assays to evaluate the migration, invasion, and proliferation of immortalized human first-trimester trophoblast cell lines
Uploaded: 2019-03-05T12:30:00
Description: Watch this Scientific Journal Video about In Vitro Assays to Evaluate the Migration, Invasion, and Proliferation of Immortalized Human First-trimester Trophoblast Cell Lines at JoVE.com

作者感谢 gil mor 博士提供了 sw.71 细胞。这项研究得到了阿尔伯特·麦肯恩·学者奖的支持。

1. 细胞制备
<ol><li>将 t-75 瓶中的细胞保持在37°c、5% co2和95% 湿度的标准生长介质中。 请注意:这些实验中使用的细胞是不朽的头三个月滋养细胞细胞系 swa. 71 (Swan.71) 和 htr-8/svneo9,10。ssw.71 细胞被维持在 dmemci-12 中, 辅以10% 的热灭活胎牛血清 (fbs), 1.0 mm 丙酮酸钠、10 mm hepes、0.10 mm mM 非必需氨基酸、100 unitsl 青霉素和100μgml 链霉素。h...

<ol>
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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Shared+and+disparate+components+of+the+pathophysiologies+of+fetal+growth+restriction+and+preeclampsia.">Shared and disparate components of the pathophysiologies of fetal growth restriction and preeclampsia.</a> American Journal of Obstetrics &amp; Gynecology. 195 (1), 40-49 (2006).</li><li>Kn&#246;fler, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Critical+growth+factors+and+signalling+pathways+controlling+human+trophoblast+invasion.">Critical growth factors and signalling pathways controlling human trophoblast invasion.</a> The International Journal of Developmental Biology. 54 (2-3), 269-280 (2010).</li><li>Huber, A. V., Saleh, L., Bauer, S., Husslein, P., Kn&#246;fler, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TNF&#x3b1;-Mediated+Induction+of+PAI-1+Restricts+Invasion+of+HTR-8/SVneo+Trophoblast+Cells.">TNF&#x3b1;-Mediated Induction of PAI-1 Restricts Invasion of HTR-8/SVneo Trophoblast Cells.</a> Placenta. 27 (2), 127-136 (2006).</li><li>Nadeem, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nodal+Signals+through+Activin+Receptor-Like+Kinase+7+to+Inhibit+Trophoblast+Migration+and+Invasion.">Nodal Signals through Activin Receptor-Like Kinase 7 to Inhibit Trophoblast Migration and Invasion.</a> The American Journal of Pathology. 178 (3), 1177-1189 (2011).</li><li>Onogi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia+inhibits+invasion+of+extravillous+trophoblast+cells+through+reduction+of+matrix+metalloproteinase+(MMP)-2+activation+in+the+early+first+trimester+of+human+pregnancy.">Hypoxia inhibits invasion of extravillous trophoblast cells through reduction of matrix metalloproteinase (MMP)-2 activation in the early first trimester of human pregnancy.</a> Placenta. 32 (9), 665-670 (2011).</li><li>Straszewski-Chavez, S. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+isolation+and+characterization+of+a+novel+telomerase+immortalized+first+trimester+trophoblast+cell+line,+Swan+71.">The isolation and characterization of a novel telomerase immortalized first trimester trophoblast cell line, Swan 71.</a> Placenta. 30 (11), 939-948 (2009).</li><li>Graham, C. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+and+characterization+of+first+trimester+human+trophoblast+cells+with+extended+lifespan.">Establishment and characterization of first trimester human trophoblast cells with extended lifespan.</a> Experimental Cell Research. 206 (2), 204-211 (1993).</li><li>Liang, C. C., Park, A. Y., Guan, 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=In+vitro+scratch+assay:+a+convenient+and+inexpensive+method+for+analysis+of+cell+migration+in+vitro.">In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro.</a> Nature Protocols. 2 (2), 329-333 (2007).</li><li>Kisanga, E. P., Tang, Z., Guller, S., Whirledge, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucocorticoid+signaling+regulates+cell+invasion+and+migration+in+the+human+first-trimester+trophoblast+cell+line+Sw.+71.">Glucocorticoid signaling regulates cell invasion and migration in the human first-trimester trophoblast cell line Sw. 71.</a> American Journal of Reproductive Immunology. , e12974 (2018).</li><li>Berthois, Y., Katzenellenbogen, J. A., Katzenellenbogen, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenol+red+in+tissue+culture+media+is+a+weak+estrogen:+implications+concerning+the+study+of+estrogen-responsive+cells+in+culture.">Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture.</a> Proceedings of the National Academy of Sciences of the United States of America. 83 (8), 2496-2500 (1986).</li><li>Cummings, B. S., Schnellmann, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cell+death+in+mammalian+cells.">Measurement of cell death in mammalian cells.</a> Current Protocols in Pharmacology. , (2004).</li></ol>

这个程序建立在使用迁移和入侵检测的基础上, 包括细胞增殖率作为一个潜在的贡献者测量的营养细胞运动的差异。一起使用, 这些体外检测是简单而有效的方法, 可用于识别调节滋养细胞迁移的细胞途径。如上所述, 滋养细胞可以用激素、细胞因子、生长因子或其他分子来治疗, 以确定它们对滋养细胞运动的影响。或者, 目标蛋白可能会从细胞中耗尽, 以阐明它们在滋养细胞运动中的功能作用。确?...

胎盘发育是建立妊娠的关键一步, 影响孕产妇和胎儿的健康。然而, 这一过程发生的机械基础并不完全了解。细胞迁移是一个重要的生物过程, 有助于怀孕期间胎盘的建立和功能。囊胚植入后, 滋养细胞分化为绒毛滋养细胞, 覆盖绒毛膜的表面, 参与气体和营养交换, 以及从绒毛中迁移出来的外生滋养细胞 (evt)并侵入母体和血管 1.evt 的迁移对于重塑产妇螺旋动脉和建立子宫胎盘循环以支持胎儿生长至关重要 2。妊娠期间滋养细胞入侵不足会导致胎盘发育异常, 并可能导致妊娠并发症, 如先兆子宫炎、妊娠期高血压、子宫内生长受限和早产3, 4. 我的工作是什么？因此, 了解影响滋养细胞运动的因素对于确定正常胎盘所需的途径至关重要。
滋养细胞迁移是由一个复杂的信号分子网络控制的, 包括生长因子、细胞因子、激素和血管生成因子5。由于在体内研究胎盘的局限性, 利用永生人类滋养细胞系进行体外检测对于确定导致滋养细胞运动的因素至关重要。划痕和入侵检测已被广泛用于定量评估单个分子在滋养细胞迁移中的作用 6,7, 8.然而, 虽然同时研究细胞入侵的变化可能会导致迁移的变化, 但这两种检测并没有考虑到可能导致细胞迁移率改变的其他机制。例如, 细胞增殖率的降低可能会导致可供迁移的细胞减少。
在这里, 我们描述了使用划痕、细胞增殖和细胞入侵检测来评估滋养细胞迁移的体外定量方法。在划痕试验中, 在细胞单层中创建一个均匀的伤口, 并通过对伤口内细胞的大小和密度进行自动延时成像来测量细胞的迁移以填补缝隙。细胞增殖分析的基础是计算每个时间点的细胞与已知的细胞起始量的比率。在细胞浸润试验中, 细胞被播种在细胞外基质包膜细胞培养插入室 (如 tranwell) 上, 并计算出通过细胞外基质侵入的细胞数量对趋化剂的反应。
划痕检测是一个简单而有效的工具, 可用于确定不同的环境条件如何影响细胞迁移。增殖和入侵检测随后可用于确定细胞增殖和入侵对细胞迁移总体变化的贡献。这些检测方法总体上提供了对生物过程的可靠测量, 这些过程可能有助于细胞的运动。在所述检测中使用了两个不朽的头三个月滋养细胞细胞系: swan.71 (Swan.71) 和 htr-8/svneo9,10。然而, 这些检测也可以优化用于其他细胞类型, 以识别细胞迁移和入侵的关键模块。

<table>
	<tbody>
		<tr>
			<td>0.4% Trypan blue stain</td>
			<td>Thermo Fisher Scientific</td>
			<td>15250-061</td>
			<td></td>
		</tr>
		<tr>
			<td>0.5% Trypsin-EDTA</td>
			<td>Life Technologies</td>
			<td>15400-054</td>
			<td></td>
		</tr>
		<tr>
			<td>1.0 M HEPES</td>
			<td>AmericanBio</td>
			<td>AB06021-00100</td>
			<td></td>
		</tr>
		<tr>
			<td>10 mM MEM non-essential amino acids</td>
			<td>Life Technologies</td>
			<td>11140-050</td>
			<td></td>
		</tr>
		<tr>
			<td>100 mM sodium pyruvate</td>
			<td>Life Technologies</td>
			<td>11360-070</td>
			<td></td>
		</tr>
		<tr>
			<td>24-well plate transwell inserts</td>
			<td>Corning</td>
			<td>3422</td>
			<td>Contain polycarbonate filters with an 8 &#956;m pore size</td>
		</tr>
		<tr>
			<td>Charcoal dextran-stripped FBS</td>
			<td>Gemini Bio-Products</td>
			<td>100-119</td>
			<td></td>
		</tr>
		<tr>
			<td>Countess II Automated Cell Counter</td>
			<td>Thermo Fisher Scientific</td>
			<td>AMQAX1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Crystal Violet</td>
			<td>Sigma Aldrich</td>
			<td>C0775</td>
			<td></td>
		</tr>
		<tr>
			<td>Cytoseal 60</td>
			<td>Thermo Fisher Scientific</td>
			<td>8310-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Dexamethasone (Dex; 1, 4-pregnadien-9&#945;-fluoro-16&#945;-methyl-11&#946;, 17, 21-triol-3, 20-dione; &#8805;98% TLC)</td>
			<td>Steraloids</td>
			<td>P0500-000</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/Ham&#8217;s F12</td>
			<td>Life Technologies</td>
			<td>11330-032</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Sigma Aldrich</td>
			<td>F2442</td>
			<td></td>
		</tr>
		<tr>
			<td>IncuCyte 24-well ImageLock Plates</td>
			<td>Essen BioScience</td>
			<td>4365</td>
			<td>If using the IncuCyte Zoom imaging system, seed cells onto IncuCyte ImageLock Plates. These plates have fiducial markers on the bottom of the plate that are used as a reference for repeat imaging in a constant field of view.</td>
		</tr>
		<tr>
			<td>IncuCyte software</td>
			<td>Essen BioScience</td>
			<td>2010A Rev2</td>
			<td></td>
		</tr>
		<tr>
			<td>IncuCyte ZOOM system</td>
			<td>Essen BioScience</td>
			<td>N/A</td>
			<td>Scratch assay images were captured and analyzed using the preset &#8220;Scratch Wound&#8221; parameters on the IncuCyte ZOOM system. Other time-lapse microscopes may also be used.</td>
		</tr>
		<tr>
			<td>Matrigel Membrane Matrix</td>
			<td>Becton Dickinson</td>
			<td>356231</td>
			<td>Growth factor reduced, phenol-red free</td>
		</tr>
		<tr>
			<td>Micro Slides</td>
			<td>VWR International, LLC</td>
			<td>48311-7003</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope cover glass</td>
			<td>Fisher Scientific</td>
			<td>12-545-E</td>
			<td></td>
		</tr>
		<tr>
			<td>Olympus IX71 inverted microscope</td>
			<td>Olympus</td>
			<td>IX71</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin (10,000 units/mL)/streptomycin (10,000 &#181;g/mL)</td>
			<td>Life Technologies</td>
			<td>151-40-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenol-red free DMEM/Ham&#39;s F12</td>
			<td>Life Technologies</td>
			<td>11039-021</td>
			<td></td>
		</tr>
		<tr>
			<td>Semi-automatic wound maker tool</td>
			<td>Essen BioScience</td>
			<td>N/A</td>
			<td></td>
		</tr>
	</tbody>
</table>

in vitro assays to evaluate the migration, invasion, and proliferation of immortalized human first-trimester trophoblast cell lines

细胞运动是胎盘发育和早孕过程中滋养细胞的重要特性。适当的滋养细胞迁移和入侵的意义表现在妊娠期疾病, 如先兆子痫和宫内生长限制, 这与不充分的滋养细胞入侵的母体血管。不幸的是, 我们对胎盘从迁移滋养细胞中发育的机制的理解是有限的。通过划痕法进行细胞迁移的体外分析是识别调节滋养细胞迁移能力的因素的有用工具。然而, 仅此检测并不能定义可能导致细胞迁移改变的细胞变化。该协议描述了三种不同的体外检测, 这是共同用于评估滋养细胞的运动: 划痕试验, 入侵试验, 和增殖试验。这里描述的协议也可以修改, 用于其他细胞系, 以量化细胞在刺激下的运动。这些方法使调查人员能够确定有助于细胞移动的个别因素, 并对细胞迁移明显变化的潜在机制进行彻底审查。

在这些实验中, 采用永生人类头三个月滋养细胞系 sw.71 和 htr-8/svneo 来确定糖皮质激素在滋养细胞迁移中的作用12。在这些实验中使用了糖皮质激素, 因为它们最近被证明可以改变滋养细胞的功能, 尽管可以使用替代治疗方法 12.在所有实验中, 用车辆 (1x pbs) 或 100 nm 的合成糖皮质激素地塞米松 (dex) 对两种细胞系进行了处理。使用自动延时?...

Watch this Scientific Journal Video about In Vitro Assays to Evaluate the Migration, Invasion, and Proliferation of Immortalized Human First-trimester Trophoblast Cell Lines at JoVE.com

In Vitro Assays to Evaluate the Migration, Invasion, and Proliferation of Immortalized Human First-trimester Trophoblast Cell Lines

在这里, 我们提出了一个高度可访问的协议, 用于评估细胞在人类滋养细胞中的运动使用三个体外检测: 划痕试验, 转井入侵试验, 细胞增殖试验。

体外检测评估神仙体头三个月滋养细胞细胞系的迁移、入侵和增殖

Cell movement is a critical property of trophoblasts during placental development and early pregnancy. The significance of proper trophoblast migration ...

这些方法可用于确定影响成虫病入侵的因素，这些因素可能提供对主要不良产科结果的洞察。该技术的优点是，它提供了可能影响成虫细胞入侵（包括增殖和迁移）的几个因素的视觉和定量分析。要开始划痕测定，将适当数量的细胞播种到24井板中，在48小时内达到100%的汇合。第二天，将介质更改为酚红色自由介质，辅以热灭活炭脱衣FBS。在划痕当天，将所需的治疗添加到每个井的介质中，以预处理细胞30分钟。要创建均匀的伤口，请将无菌 10 微升移液器尖端放在伤口制造商工具的销上。将刀尖放入第一排井中，沿伤口制造商的轨道移动板，直到移液器尖端到达油井的另一侧。然后将板移回起始位置，以确保在井的直径上不断缠绕。将尖端放入下一排油井中，然后重复，直到所有油井都刮伤。在显微镜下观察板材，确认划痕覆盖了井的宽度。接下来，小心吸气介质，用1x PBS轻轻清洗两次细胞。最后洗涤后，添加补充的酚红色无剥离或低FBS介质与所需的治疗。接下来，将带划痕孔的板放在自动延时成像器中，该图像器中含有 37 摄氏度的温度和 95% 湿度的 95% 湿度环境中的 5% 二氧化碳。根据需要定期对细胞进行成像，以便细胞完成伤口愈合。开始入侵测定，在95%湿度的大气中保持37摄氏度的温度和5%的二氧化碳的培养箱中，将适当数量的细胞播种到六孔板中。允许细胞在48小时内达到90%的汇合。第二天，将介质更改为酚红色自由介质，辅以热灭活炭脱衣FBS。将每毫升10毫克的细胞外基瓶放在冰箱的冰上，让一夜之间解冻。后天，使用冷冻移液器提示，用酚红色自由血清基质稀释每毫升10毫克的细胞外基质，最终浓度为每毫升5毫克。然后将稀释的细胞外基质的100毫升加入冷藏的24井板中。在37摄氏度下孵育板，使基质变硬。要为入侵测定准备细胞，请用1毫升1x PBS清洗细胞。然后吸气PBS，并添加500微升1倍的尝试素EDTA到每一个井。将六井板放在培养箱中，在95%的湿度环境中保持37摄氏度的温度和5%的二氧化碳，直到所有细胞都从板底分离。一旦细胞分离，加入500微升的酚红色自由血清无素介质到每一个井的三辛化细胞。然后将1000微升分离细胞转移到微离心管中，在4摄氏度下以400倍g下旋转5分钟。接下来，吸气介质，重新暂停苯酚红色自由血清基中的细胞。使用自动细胞计数器计算细胞，并调整细胞悬浮量，最终浓度为每毫升 0.5 百万细胞。在 24 井板的每个井中加入 600 微升的完整生长介质，并将预涂层刀片放入其中。然后在每个预涂层细胞插入物上添加200微升细胞，总共10万个细胞。将24井板放在培养箱中，在95%的湿度环境中保持37摄氏度的温度和5%的二氧化碳，24小时。经过夜间孵育后，从上腔和下腔中吸入剩余的介质，而不会干扰细胞外基质。然后用棉签小心地从刀片的上部取出细胞外基质和非侵入细胞。将 1x PBS 添加到井的上部和下腔，将每个刀片洗涤三次。每次洗涤后吸气 PBS。要修复连接到刀片的细胞，请将插入物放入 100%冰冷甲醇中，在 4 摄氏度下放置 30 分钟。用 1x PBS 清洗刀片三次，并在最后洗涤后取出 PBS。用0.2%水晶紫罗兰在20%乙醇中染色附着在刀片上的细胞10分钟。用去水冲洗三次，最后洗涤后吸气脱水水。空气干燥插件在室温下一小时。最后，使用钳子和剃刀刀片，小心地切割刀片的底部膜，并将其安装在玻璃滑梯上，底部朝上。使用具有 20 倍目标的光学显微镜，每个样本至少获得四个入侵细胞的独特图像。要开始增殖检测，请使用自动细胞计数器从烧瓶中计数多头细胞。在标准生长介质中，以每孔0.20万个细胞的密度将细胞播种到六孔板中。然后将细胞放在培养箱中，在95%的湿度大气中保持37摄氏度的温度和5%的二氧化碳，维持4个小时，或直到细胞粘附。接下来，将介质更改为酚红色无源介质，辅以热灭活炭脱精脱衣 FBS 和所需处理。将六井板放在培养箱中，在95%的湿度环境中保持37摄氏度的温度和5%的二氧化碳。然后更换新介质，每 48 小时辅以所需的治疗。经过24、48或72小时的治疗，用1毫升PBS清洗细胞，并在每井中加入500微升的三辛EDTA。在培养箱中放置一个六井板，直到细胞与板分离。一旦细胞从板分离，加入500微升的补充剂酚红色自由介质，以停用尝试性。最后在单独的管子中将五微升的三辛化细胞加入 Trypan Blue 的五微升中，然后通过移液混合。使用自动单元格计数器对重复的每个井中的单元格进行计数。不朽的人类前三个月成河天鹅。71个细胞在零、8和18小时内用车辆1x PBS或100微摩尔的合成糖皮质激素二甲酮在划痕测定中治疗。测量天鹅的伤口大小和伤口密度。71 和 HTR-8/SVneo 细胞用车辆或 100 纳米摩尔 dex 治疗表明，dex 治疗降低了伤口闭合率，表明糖皮质激素可以抑制细胞迁移。入侵测定后，糖皮质激素治疗降低了细胞侵入性，如入侵天鹅数量减少15%。。71 和 HTR-8/SVneo 细胞与车辆处理的细胞相比，用 100 纳米摩尔 dex 处理 24 小时。在细胞增殖分析后，糖皮质激素治疗减少了细胞增殖，如天鹅较少所示。71 和 HTR-8/SVneo 细胞处理 100 纳米摩尔 dex 与车辆处理的细胞相比。评估细胞死亡的方法可以与本程序结合使用，以确定凋亡或坏死是否有助于改变成虫细胞功能。我们独特地采用了癌症生物学中常用的标准技术来研究成虫细胞功能。没有与这些方法相关的危险。观众唯一应该采取的预防措施是使用无菌技术进行组织培养。

Research

JoVE Journal

Developmental Biology

Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays

Production and Use of Lentivirus to Selectively Transduce Primary Oligodendrocyte Precursor Cells for In Vitro Myelination Assays

生产和使用慢病毒的有选择性地转导原少突胶质前体细胞<em&gt;在体外</em&gt;髓鞘测定

Myelination is a complex process that involves both neurons and the myelin forming glial cells, oligodendrocytes in the central nervous system (CNS) and ...

科研

发育生物学

Assessment of Maternal Vascular Remodeling During Pregnancy in the Mouse Uterus

产妇血管重构的评估在怀孕期间在小鼠子宫

The placenta mediates the exchange of factors such as gases and nutrients between mother and fetus and has specific demands for supply of blood from the ...

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation

建立基因组编辑的人多能干细胞系：从定位到隔离

Genome-editing of human pluripotent stem cells (hPSCs) provides a genetically controlled and clinically relevant platform from which to understand human ...

Initiating Differentiation in Immortalized Multipotent Otic Progenitor Cells

Use of human induced pluripotent stem cells (iPSC) or embryonic stem cells (ESC) for cell replacement therapies holds great promise. Several limitations ...

Development of an In Vitro Assay to Evaluate Contractile Function of Mesenchymal Cells that Underwent Epithelial-Mesenchymal Transition

Development of an In Vitro Assay to Evaluate Contractile Function of Mesenchymal Cells that Underwent Epithelial-Mesenchymal Transition

的发展<em&gt;体外</em&gt;检测以确定间充质细胞将经历上皮 - 间质转化的收缩功能

Fibrosis is often involved in the pathogenesis of various chronic progressive diseases such as interstitial pulmonary disease. Pathological hallmark is ...

Human Primary Trophoblast Cell Culture Model to Study the Protective Effects of Melatonin Against Hypoxia/reoxygenation-induced Disruption

人胎盘滋养层细胞培养模型，研究褪黑素对缺氧的保护作用/复氧引起的中断

This protocol describes how villous cytotrophoblast cells are isolated from placentas at term by successive enzymatic digestions, followed by density ...

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

5-mC点印迹测定定量软骨细胞去分化的DNA甲基化水平<em&gt;体外</em

The dedifferentiation of hyaline chondrocytes into fibroblastic chondrocytes often accompanies monolayer expansion of chondrocytes in vitro. The global ...

Live Imaging of Mouse Secondary Palate Fusion

小鼠第二腭融合的实时成像

The fusion of the secondary palatal shelves to form the intact secondary palate is a key process in mammalian development and its disruption can lead to ...

Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea

Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea

角膜组织工程: 人角膜间质神经相互作用的体外模型

Tissue engineering has gained substantial recognition due to the high demand for human cornea replacements with an estimated 10 million people worldwide ...

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

Cell Aggregation Assays to Evaluate the Binding of the Drosophila Notch with Trans-Ligands and its Inhibition by Cis-Ligands

细胞聚集法评价果蝇切口与反式配体的结合及其对顺式配体的抑制作用

Notch signaling is an evolutionarily conserved cell-cell communication system used broadly in animal development and adult maintenance. Interaction of the ...