Name: application of aorta-gonad-mesonephros explant culture system in developmental hematopoiesis
Uploaded: 2017-11-03T13:00:00
Description: Watch this Scientific Journal Video about Application of Aorta-gonad-mesonephros Explant Culture System in Developmental Hematopoiesis at JoVE.com

我们感谢普华永在图准备方面的帮助。这项工作得到了中国国家自然科学基金 (81530004、31425016) 和中国科学技术部 (2016YFA0100500) 的资助。. 进行了实验并草拟了手稿;--编辑了手稿。两位作者阅读并批准了最后的手稿。

包括动物学科在内的所有程序都已获得中国科学院动物研究所伦理审查委员会的批准. 
 1. 材料准备 <ol> <li> 对0.65 和 #181 进行消毒; m 过滤器在紫外线灯下产生紫外线, 在清洁的长凳上为4小时, 并且在 2 h 标记以后转动滤光器。
	注意: 当使用 UV 光时, 请穿戴适当的保护. </li>
	<li> 121 和 #176 用高压灭菌器消毒不锈钢网格, 30 分钟. 
	注: 需要一定高度的不锈钢网格, 以支持气接...

<ol>
	<li>Bertrand, J. Y., 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=Haematopoietic+stem+cells+derive+directly+from+aortic+endothelium+during+development.">Haematopoietic stem cells derive directly from aortic endothelium during development.</a> Nature. 464, 108-111 (2010).</li><li>Boisset, J. C., Robin, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+the+founder+of+adult+hematopoiesis+in+the+mouse+embryo+aorta.">Imaging the founder of adult hematopoiesis in the mouse embryo aorta.</a> Cell cycle. 9, 2489-2490 (2010).</li><li>Kissa, K., Herbomel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+stem+cells+emerge+from+aortic+endothelium+by+a+novel+type+of+cell+transition.">Blood stem cells emerge from aortic endothelium by a novel type of cell transition.</a> Nature. 464, 112-115 (2010).</li><li>Zovein, A. C., 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=Fate+tracing+reveals+the+endothelial+origin+of+hematopoietic+stem+cells.">Fate tracing reveals the endothelial origin of hematopoietic stem cells.</a> Cell Stem Cell. 3, 625-636 (2008).</li><li>Kumaravelu, P., 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=Quantitative+developmental+anatomy+of+definitive+haematopoietic+stem+cells+long-term+repopulating+units+(HSC/RUs):+role+of+the+aorta-gonad-mesonephros+(AGM)+region+and+the+yolk+sac+in+colonisation+of+the+mouse+embryonic+liver.">Quantitative developmental anatomy of definitive haematopoietic stem cells long-term repopulating units (HSC/RUs): role of the aorta-gonad-mesonephros (AGM) region and the yolk sac in colonisation of the mouse embryonic liver.</a> Development. 129, 4891-4899 (2002).</li><li>Orkin, S. H., Zon, L. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hematopoiesis:+an+evolving+paradigm+for+stem+cell+biology.">Hematopoiesis: an evolving paradigm for stem cell biology.</a> Cell. 132, 631-644 (2008).</li><li>Medvinsky, A., Dzierzak, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Definitive+hematopoiesis+is+autonomously+initiated+by+the+AGM+region.">Definitive hematopoiesis is autonomously initiated by the AGM region.</a> Cell. 86, 897-906 (1996).</li><li>Peeters, M., 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=Ventral+embryonic+tissues+and+Hedgehog+proteins+induce+early+AGM+hematopoietic+stem+cell+development.">Ventral embryonic tissues and Hedgehog proteins induce early AGM hematopoietic stem cell development.</a> Development. 136, 2613-2621 (2009).</li><li>Taoudi, S., Medvinsky, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+identification+of+the+hematopoietic+stem+cell+niche+in+the+ventral+domain+of+the+embryonic+dorsal+aorta.">Functional identification of the hematopoietic stem cell niche in the ventral domain of the embryonic dorsal aorta.</a> PNAS. 104, 9399-9403 (2007).</li><li>Crisan, M., 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=BMP+and+Hedgehog+Regulate+Distinct+AGM+Hematopoietic+Stem+Cells+Ex+Vivo.">BMP and Hedgehog Regulate Distinct AGM Hematopoietic Stem Cells Ex Vivo.</a> Stem Cell Reports. 6, 383-395 (2016).</li><li>Durand, C., 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=Embryonic+stromal+clones+reveal+developmental+regulators+of+definitive+hematopoietic+stem+cells.">Embryonic stromal clones reveal developmental regulators of definitive hematopoietic stem cells.</a> PNAS. 104, 20838-20843 (2007).</li><li>Fleury, M., Petit-Cocault, L., Clay, D., Souyri, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mpl+receptor+defect+leads+to+earlier+appearance+of+hematopoietic+cells/hematopoietic+stem+cells+in+the+Aorta-Gonad-Mesonephros+region,+with+increased+apoptosis.">Mpl receptor defect leads to earlier appearance of hematopoietic cells/hematopoietic stem cells in the Aorta-Gonad-Mesonephros region, with increased apoptosis.</a> Int J Dev Biol. 54, 1067-1074 (2010).</li><li>Lv, J., Wang, L., Gao, Y., Ding, Y. Q., Liu, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=5-hydroxytryptamine+synthesized+in+the+aorta-gonad-mesonephros+regulates+hematopoietic+stem+and+progenitor+cell+survival.">5-hydroxytryptamine synthesized in the aorta-gonad-mesonephros regulates hematopoietic stem and progenitor cell survival.</a> J Exp Med. 214, 529-545 (2017).</li><li>Souilhol, C., 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=Inductive+interactions+mediated+by+interplay+of+asymmetric+signalling+underlie+development+of+adult+haematopoietic+stem+cells.">Inductive interactions mediated by interplay of asymmetric signalling underlie development of adult haematopoietic stem cells.</a> Nature communications. 7, 10784 (2016).</li><li>Fitch, S. R., 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=Signaling+from+the+sympathetic+nervous+system+regulates+hematopoietic+stem+cell+emergence+during+embryogenesis.">Signaling from the sympathetic nervous system regulates hematopoietic stem cell emergence during embryogenesis.</a> Cell Stem Cell. 11, 554-566 (2012).</li><li>Bianchi, M., Moser, C., Lazzarini, C., Vecchiato, E., Crespi, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forced+swimming+test+and+fluoxetine+treatment:+in+vivo+evidence+that+peripheral+5-HT+in+rat+platelet-rich+plasma+mirrors+cerebral+extracellular+5-HT+levels,+whilst+5-HT+in+isolated+platelets+mirrors+neuronal+5-HT+changes.">Forced swimming test and fluoxetine treatment: in vivo evidence that peripheral 5-HT in rat platelet-rich plasma mirrors cerebral extracellular 5-HT levels, whilst 5-HT in isolated platelets mirrors neuronal 5-HT changes.</a> Exp Brain Res. 143, 191-197 (2002).</li><li>Ortiz, J., Artigas, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+monoamine+uptake+inhibitors+on+extracellular+and+platelet+5-hydroxytryptamine+in+rat+blood:+different+effects+of+clomipramine+and+fluoxetine.">Effects of monoamine uptake inhibitors on extracellular and platelet 5-hydroxytryptamine in rat blood: different effects of clomipramine and fluoxetine.</a> Br J Pharmacol. 105, 941-946 (1992).</li><li>Wong, D. 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众所周知, 成熟的干细胞在骨髓中可以重新填充受照射的接受者的血液系统。与这些功能干细胞不同的是, 在小鼠胚胎的周年大会上出现的干细胞是不成熟的。直接移植结果表明, I 型 (ve cad+ CD45- CD41+) 和类型 II (ve cad+ CD45+) 前干细胞具有没有重构能力20。利用股东大会的外植体培养体系, 在干细胞的新生中, 可以在移植试验中重建受体<sup clas...

造血干细胞 (干细胞) 是组织特异的成体干细胞, 具有多向的潜能, 包括红、髓细胞和淋巴细胞, 以及自我的能力。最近的研究表明, 最早的干细胞产生于一个专门的内皮细胞, 称为 hemogenic 内皮细胞 (他), 通过内皮到造血转移 (EHT) 在腹壁的背主动脉1,2 ,3,4。一旦从胚胎 (E) 10.5 到 E12.5 在小鼠胚胎中形成了主动脉-性腺-肾 (年会) 区域, 干细胞就会迁移到胎儿肝脏进行扩张, 最终在个体的生命中殖民骨髓以维持成人造血。5,6. 尽管这一研究已经进行了许多年, 但 HSC 产生和发展的基本机制仍未得到完全理解。
与体外不同的是, 斑马鱼胚胎的受精和发育, 小鼠胚胎的宫内发育使得在胚胎发生过程中的最终造血研究更不方便。虽然使用击倒 (ko) 小鼠的基因实验是常用的, 但某些高鼠的缺乏也限制了它们在造血研究领域的应用。此外,在体内抢救实验在 KO 小鼠中不易进行。自1996年以来, 在该领域的先驱在7中为造血研究开发了 "年度" 外植体培养。在这一文化体系的帮助下, 已经确定了股东大会区域的腹侧组织以促进 HSC 活动, 而背部组织则发挥相反的作用8,9。在 HSC 的发展中, 应用了 Mpl、蛋白、蛋白、BMP 和刺猬的信号, 确定了血清素、外植体培养系统的作用10、11、12、13、14. 重要的是, 它也是一种常用的方法来挽救突变体胚胎的造血缺损13,15。

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		<tr height="21" style="height:15.75pt">
			<td class="xl67" height="21" style="width:162pt;height:15.75pt" width="216">durapore 0.65 &#181;m filters</td>
			<td class="xl68" style="width:81pt" width="108">Millipore</td>
			<td class="xl69" style="width:89pt" width="119">R5BA63787</td>
			<td class="xl67" style="width:125pt" width="167">AGM explant culture</td>
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		<tr height="42" style="height:31.5pt">
			<td class="xl67" height="42" style="height:31.5pt">M5300 long-term culture medium</td>
			<td class="xl68" style="width:81pt" width="108">Stem Cell Technologies</td>
			<td class="xl69" style="width:89pt" width="119">5300</td>
			<td class="xl67">AGM explant culture</td>
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			<td class="xl68" height="21" style="width:162pt;height:15.75pt" width="216">Trizol&#160;</td>
			<td class="xl68" style="width:81pt" width="108">Tiangen</td>
			<td class="xl69" style="width:89pt" width="119">5829</td>
			<td class="xl67">RNA extration</td>
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			<td class="xl68" height="21" style="width:162pt;height:15.75pt" width="216">M-MLV reverse transcriptase</td>
			<td class="xl68" style="width:81pt" width="108">Promega</td>
			<td class="xl69" style="width:89pt" width="119">90694</td>
			<td class="xl67">reverse transcription</td>
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			<td class="xl68" height="21" style="width:162pt;height:15.75pt" width="216">SYBR Green</td>
			<td class="xl68" style="width:81pt" width="108">Qiagen</td>
			<td class="xl69" style="width:89pt" width="119">A6002</td>
			<td class="xl67">quantatitive real-time PCR</td>
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		<tr height="21" style="height:15.75pt">
			<td class="xl68" height="21" style="width:162pt;height:15.75pt" width="216">protease inhibitor</td>
			<td class="xl68" style="width:81pt" width="108">Roche</td>
			<td class="xl69" style="width:89pt" width="119">55622</td>
			<td class="xl67">Protein extration</td>
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		<tr height="21" style="height:15.75pt">
			<td class="xl68" height="21" style="width:162pt;height:15.75pt" width="216">collagenase</td>
			<td class="xl68" style="width:81pt" width="108">Sigma</td>
			<td class="xl69" style="width:89pt" width="119">C2674</td>
			<td class="xl67">digestion of AGM tissues</td>
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		<tr height="42" style="height:31.5pt">
			<td class="xl68" height="42" style="width:162pt;height:31.5pt" width="216">MethoCult GF M3434 medium</td>
			<td class="xl68" style="width:81pt" width="108">Stem Cell Technologies</td>
			<td class="xl69" style="width:89pt" width="119">3434</td>
			<td class="xl67">CFU-C assay</td>
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		<tr height="42" style="height:31.5pt">
			<td class="xl68" height="42" style="width:162pt;height:31.5pt" width="216">ultra-low attachment 24-well plates</td>
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			<td class="xl69" style="width:89pt" width="119">3473</td>
			<td class="xl67">CFU-C assay</td>
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			<td class="xl67" height="63" style="height:47.25pt">C57BL/6 CD45.2 mice</td>
			<td class="xl68" style="width:81pt" width="108">Beijing HFK Bioscience Co. Ltd</td>
			<td class="xl68" style="width:89pt" width="119"></td>
			<td class="xl67">CFU-S assay</td>
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		<tr height="63" style="height:47.25pt">
			<td class="xl68" height="63" style="width:162pt;height:47.25pt" width="216">C57BL/6 CD45.1 mice</td>
			<td class="xl68" style="width:81pt" width="108">Beijing HFK Bioscience Co. Ltd</td>
			<td class="xl68" style="width:89pt" width="119"></td>
			<td class="xl67">Long-term transplantation</td>
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		<tr height="42" style="height:31.5pt">
			<td class="xl68" height="42" style="width:162pt;height:31.5pt" width="216">phosphate buffered saline</td>
			<td class="xl68" style="width:81pt" width="108">Life Technologines</td>
			<td class="xl69" style="width:89pt" width="119">8115284</td>
			<td class="xl68" style="width:125pt" width="167">AGM Collection</td>
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			<td class="xl68" height="21" style="width:162pt;height:15.75pt" width="216">Penicillin-Streptomycin solution</td>
			<td class="xl68" style="width:81pt" width="108">HyClone</td>
			<td class="xl68" style="width:89pt" width="119">SV30010</td>
			<td class="xl68" style="width:125pt" width="167">AGM explant culture</td>
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		<tr height="42" style="height:31.5pt">
			<td class="xl68" height="42" style="width:162pt;height:31.5pt" width="216">anti-CD45.2-PE-CY7</td>
			<td class="xl68" style="width:81pt" width="108">eBioscience</td>
			<td class="xl68" style="width:89pt" width="119">25-0454-80</td>
			<td class="xl68" style="width:125pt" width="167">Long-term transplantation</td>
		</tr>
		<tr height="42" style="height:31.5pt">
			<td class="xl68" height="42" style="width:162pt;height:31.5pt" width="216">anti-CD45-FITC</td>
			<td class="xl68" style="width:81pt" width="108">eBioscience</td>
			<td class="xl68" style="width:89pt" width="119">11-0451-81</td>
			<td class="xl68" style="width:125pt" width="167">Long-term transplantation</td>
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		<tr height="105" style="height:78.75pt">
			<td class="xl68" height="105" style="width:162pt;height:78.75pt" width="216">UV lamp</td>
			<td class="xl68" style="width:81pt" width="108">Beijing jiangmorning yuan bio-technology co., LTD</td>
			<td class="xl68" style="width:89pt" width="119">039-14402</td>
			<td class="xl68" style="width:125pt" width="167">power: 30W</td>
		</tr>
		<tr height="63" style="height:47.25pt">
			<td class="xl68" height="63" style="width:162pt;height:47.25pt" width="216">stainless steel mesh</td>
			<td class="xl68" style="width:81pt" width="108">AS ONE SHANGHAI Corporation</td>
			<td class="xl68" style="width:89pt" width="119">2-9817-10&#160;</td>
			<td class="xl68" style="width:125pt" width="167">aperture diameter: 2.5 mm</td>
		</tr>
		<tr height="42" style="height:31.5pt">
			<td class="xl68" height="42" style="width:162pt;height:31.5pt" width="216">hydrocortisone</td>
			<td class="xl68" style="width:81pt" width="108">Sigma</td>
			<td class="xl68" style="width:89pt" width="119">H0396</td>
			<td class="xl68" style="width:125pt" width="167">selectively diluted into M5300 medium</td>
		</tr>
	</tbody>
</table>

application of aorta-gonad-mesonephros explant culture system in developmental hematopoiesis

使用小鼠胚胎进行造血研究的局限性是在手术中增加了不便, 这主要是由于胚胎的宫内发育。尽管来自击倒 (ko) 小鼠的基因数据令人信服, 但在需要时为所有基因生成高鼠并不现实。此外, 执行在体内抢救实验, 以巩固从 KO 小鼠获得的数据是不方便的。为了克服这些局限性, 主动脉-性腺-肾 (年会) 外植体培养作为研究造血干细胞 (HSC) 发育的适宜系统而发展起来。特别是在抢救实验中, 可用于恢复高鼠的造血损伤。通过在培养基中加入适当的化学物质, 可以重新激活受损的信号, 或抑制上调的通路。利用这种方法, 可以进行大量的实验, 以确定 hsc 发展的关键调控因子, 包括 hsc 相关基因在 mRNA 和蛋白质水平、菌落形成能力和重构能力上的表达。这一系列的实验将有助于确定哺乳动物 HSC 发育所必需的基本机制。

最近发表的一篇报道说, 内皮细胞衍生的血清素通过抑制干细胞的亡途径, 促进了其生存.13。为了证实血清素对 HSC 发育的促进作用, 氟西汀被列入。作为一种选择性血清素再抑制剂 (SSRI), 氟西汀已被证明抑制血清素吸收在周围组织16,17,18。采用上述方法对氟西汀对 HSC 培养的影响进行了...

Watch this Scientific Journal Video about Application of Aorta-gonad-mesonephros Explant Culture System in Developmental Hematopoiesis at JoVE.com

Application of Aorta-gonad-mesonephros Explant Culture System in Developmental Hematopoiesis

本议定书描述使用培养的主动脉-性腺-肾的表达分析, 菌落形成单位在文化和脾脏, 并长期重建, 以确定影响的调节因子和信号通路的造血干细胞发育。这已被证明是一个有效的系统研究造血干细胞生物学和功能。

主动脉-性腺-肾外植体培养体系在发育性造血中的应用

The limitation of using mouse embryos for hematopoiesis studies is the added inconvenience in operations, which is largely due to the intrauterine ...

The overall goal of this experiment is to use an aorta-gonad-mesonephros explant culture to investigate the critical regulators of hematopoietic stem cell development. This method can help answer key questions in HSC biology and function fields about how to identify the factors involved in the regulation of HSC development in mammals. The main advantage of this technique is that these regulators can be verified through DNA expression analysis and the colony formation ability and the reconstitution capacity of the HSC.Generally individuals new to this method will struggle because the precise separation of AGM from the surrounding tissues in the trunk region requires practice to master. Before beginning the procedure place stainless steel meshes into individual wells of a six-well plate containing two milliliters of freshly prepared fluoxetine-supplemented medium per well. Place one UV-sterilized 0.65 microliter filter per experimental well into a glass cell culture dish containing fresh boiling water for two three-minute washes, immersing the filters in a container of room temperature PBS for two minutes after each wash.Then place one filter onto each steel mesh at the air-liquid interface of each well. Next use the dissecting scissors to strip the uterus from an embryonic day-11 pregnant mouse and harvest the embryos from the uterus. Using dissecting scissors remove the yolk sac, umbilical cord and viscera from the embryo body.And carefully remove the dorsal nerve tissues and surrounding muscular tissues. Using forceps remove the tissues from the anterior and hind limbs. And separate the AGM region from the embryo body.Then place each AGM onto a single filter at the air-liquid interface of each well. Place the cultures in a 37 degree Celsius and 5%CO2 incubator. After two to three days, transfer the AGM samples into individual 1.5-milliliter microcentrifuge tubes containing 200 microliters of collagenase for their dissociation at 37 degrees Celsius for about 20 minutes with shaking every five minutes.At the end of the digestion stop the reactions with 200 microlilters of serum per explant. Transfer the entire contents of each tube into individual wells of a 24-well ultra-low attachment plate containing 500 to 600 microliters of M3434 medium per well. Mix the cells in each well with a one milliliter syringe without a needle.Place the plate in the cell culture incubator. The number of colony-forming units within each well can then be scored after seven to 10 days of culture. For in vivo AGM explant transplantation after two to three days of AGM culture as just demonstrated, lethally eradiate eight to 10-week-old male C57 black six mice with nine grays of radiation.Four to five hours later dissociate the harvested explants in 200 microliters of collagenase per samples as demonstrated. Load of 0.5 to one embryo equivalent of the resulting single-cell suspensions into one one-milliliter syringe per recipient animal. Place the first irradiated mouse into a restrainer and wipe the tail with a 75%ethanol-soaked cotton swab to promote tail vein vasodilation.Intravenously inject the entire contents of one syringe into a dilated tail vein. Use an antiseptic swab to stop the bleeding after removal of the needle. Then return the animal to its cage, and inject the next animal.11 days after the adoptive transfer the spleens can be fixed in Bouin's solution for macroscopic enumeration of the visible spleen colonies. Fluoxetine-supplemented AGM cultures demonstrate an increased expression of Runx1, a pivotal gene for HSC development at the mRNA and protein expression levels. Fluoxetine can also significantly increase the colony formation ability of HSC in the AGM including burst-forming unit erythroids, colony-forming unit granular monocytes and colony-forming unit granulocytes, erythrocytes, macrophages and megakaryocytes.Further in vivo transplantation of fluoxetine-treated HSC isolated from AGM explants results in increased numbers of colonies in the spleens of irradiated adult recipient animals. After watching this video, you should have a good understanding of how to isolate the AGM and to perform an AGM explant culture for the identification of the critical regulators of HSC development within the AGM.

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

Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol

从人多能干细胞使用高度可重复的小分子为基础的分化协议心肌细胞规模化生产

Maximizing the benefit of human pluripotent stem cells (hPSCs) for research, disease modeling, pharmaceutical and clinical applications requires robust ...

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

Application of RNAi and Heat-shock-induced Transcription Factor Expression to Reprogram Germ Cells to Neurons in C. elegans

rna 干扰和热休克诱导转录因子表达在重编程生殖细胞到神经元中的应用C. 线虫

Studying the cell biological processes during converting the identities of specific cell types provides important insights into mechanism that maintain ...

Efficient Generation of Pancreas/Duodenum Homeobox Protein 1+ Posterior Foregut/Pancreatic Progenitors from hPSCs in Adhesion Cultures

Efficient Generation of Pancreas/Duodenum Homeobox Protein 1+ Posterior Foregut/Pancreatic Progenitors from hPSCs in Adhesion Cultures

高效生成粘附培养中 Hpsc 的胰腺-二叶多布蛋白 1+后后福格内胰腺祖细胞

Human pluripotent stem cell (hPSC)-derived pancreatic cells are a promising cell source for regenerative medicine and a platform to study human ...

In Vitro Generation of Somite Derivatives from Human Induced Pluripotent Stem Cells

人诱导多能干细胞体外生成体细胞衍生物的研究

In response to signals such as WNTs, bone morphogenetic proteins (BMPs), and sonic hedgehog (SHH) secreted from surrounding tissues, somites (SMs) give ...

Hemogenic Endothelium Differentiation from Human Pluripotent Stem Cells in A Feeder- and Xeno-free Defined Condition

在无进料和异种定义的条件下,从人类多能干细胞中血性内皮物分化

Deriving functional hematopoietic stem and progenitor cells (HSPCs) from human pluripotent stem cells (PSCs) have been an elusive goal for autologous ...

In vitro Neuromuscular Junction Induced from Human Induced Pluripotent Stem Cells

从人类诱导多能干细胞诱导的体外神经肌肉结点

The neuromuscular junction (NMJ) is a specialized synapse that transmits action potentials from the motor neuron to skeletal muscle for mechanical ...

A 3-D Tail Explant Culture to Study Vertebrate Segmentation in Zebrafish

研究斑马鱼脊椎动物分割的三维尾部外植培养

Vertebrate embryos pattern their major body axis as repetitive somites, the precursors of vertebrae, muscle, and skin. Somites progressively segment from ...