Name: studying cryptosporidium infection in 3d tissue-derived human organoid culture systems by microinjection
Uploaded: 2019-09-14T13:00:00
Description: Watch this Scientific Journal Video about Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection at JoVE.com

我们感谢美国亚利桑那州亚利桑那大学农业和生命科学学院动物和比较生物医学学院的Deborah A. Schaefer帮助我们进行卵母细胞生产和分析。我们还感谢弗朗切斯基显微镜和成像中心和D.L.Mullendore在华盛顿州立大学的TEM准备和成像的分离器官卵母细胞。
D.D. 是荷兰科学研究组织（NWO-ALW， 016.Veni.171.015） 的 VENI 赠款的接受者。I.H. 是荷兰科学研究组织（NWO-ALW，863.14.002）的 VENI 赠款的接受者，并得到欧盟委员会玛丽·居里研究金（提案 330571 FP7-PEOPLE-2012-IIF）的支持。根据ERC高级赠款协议第67013号，从欧洲研究理事会获得第67013号高级赠款协议，从NIH NIAIH获得R21 AT009174至RMO的资助。这项工作是Oncode研究所的一部分，该研究所部分由荷兰癌症协会资助，由荷兰癌症协会的赠款资助。

所有组织处理和切除均在患者同意下根据机构审查委员会 （IRB） 批准的协议进行。
1. 制备用于注射的C.帕武姆卵囊
注：隐孢子虫是从商业来源购买的（见材料表）。这些卵母细胞在小牛中生产，并储存在磷酸盐缓冲盐水（PBS）与抗生素。它们可在4°C下储存约3个月，切勿冷冻。我们通常在一个月内使用卵母细胞。有?...

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肠道和气道类有机物中隐孢子虫寄生虫的培养为研究宿主-寄生虫相互作用提供了一个精确的模型，但还有许多其他应用。例如，目前选择和传播转基因隐孢子虫寄生虫的方法要求在小鼠19中通过，这不允许分离对体内感染至关重要的寄生虫。隐孢子虫的有机体培养提供了这个程序的替代方案。然而，我们注意到，电孔孢子团聚集在一起，并?...

治疗和预防隐孢子虫感染的药物或疫苗的开发一直受到缺乏体外系统，精确模仿人体体内情况1，2的阻碍。许多目前可用的系统要么只允许短期感染（<5天），要么不支持寄生虫3，4的完整生命周期。其他能够完全发展寄生虫的系统是基于不朽的细胞系或癌细胞系，它们不能忠实地概括人类的生理状况5，6，7.有机体或"小器官"是3D组织衍生结构，生长在细胞外基质中，辅以各种组织特定的生长因子。有机体是从各种器官和组织发展起来的。它们具有遗传稳定性，可概括其来源器官的大部分功能，可在培养中长期保存。我们开发了一种用隐孢子虫感染人类肠道和肺器官的方法，为研究与肠道和呼吸隐孢子虫病相关的宿主-寄生虫相互作用提供了精确的体外模型8 ，9，10，11，12，13。与其他已发表的培养模型相比，有机体系统代表现实生活中的宿主寄生虫相互作用，允许完成生命周期，以便研究寄生虫生命周期的所有阶段，并保持寄生虫繁殖长达28天10。
隐孢子虫是一种感染呼吸道和肠道上皮的寄生虫，引起长期腹泻病。耐抗环境阶段是卵母细胞，发现在受污染的食物和水14。一旦摄入或吸入，卵囊肿和释放四个附着在上皮细胞的孢子石。孢子石在宿主细胞上滑行，并接合宿主细胞受体，但寄生虫没有完全侵入细胞，似乎诱导宿主细胞吞没它15。寄生虫在细胞内但细胞外质室内化，留在细胞的表层，在寄生性腹腔内复制。它经历了两轮无性繁殖——这个过程叫做美罗戈尼。在美罗戈尼期间，I型美龙特人发育，其中含有八种甲子，它们被释放以侵入新的细胞。这些美罗佐石侵入新的细胞，发展成含有四个美罗索石II型的梅龙特。这些美罗佐特，当被释放，感染细胞，并发展成宏加蒙特和微加蒙特。微配因子被释放，使大玩家受精，产生成熟成卵囊的酶。成熟的卵母细胞随后被释放到流明中。卵囊是薄壁，立即排泄重新感染上皮，或厚壁被释放到环境中，以感染下一个主机14。隐孢子虫生命周期的所有阶段都已在我们的第10组先前开发的有机体培养系统中被确定。
由于人体器官忠实地复制人体组织9，11，13，并支持隐孢子虫10的所有复制阶段，它们是理想的组织培养系统研究隐孢子虫生物学和宿主寄生虫相互作用。在这里，我们描述了感染有机体与隐孢子虫和排泄孢子虫虫的程序，并分离在这个组织培养系统中产生的新卵母细胞。

<table>
<tbody>
<tr>
<td>Basement membrane extract (extracellular matrix)</td>
<td>amsbio</td>
<td>3533-010-02</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Crypt-a-Glo antibody (Oocyst specific antibody)</td>
<td>Waterborne, Inc</td>
<td>A400FLR-1X</td>
<td>Final Concentration = Use 2-3 drops/slide</td>
</tr>
<tr>
<td>Crypto-Grab IgM coated Magnetic beads</td>
<td>Waterborne, Inc</td>
<td>IMS400-20</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Dynamag 15 rack</td>
<td>Thermofisher Scientific</td>
<td>12301D</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Dynamag 2 rack</td>
<td>Thermofisher Scientific</td>
<td>12321D</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>EMD Millipore&nbsp;Isopore Polycarbonate Membrane Filters- 3&micro;m</td>
<td>EMD-Millipore</td>
<td>TSTP02500</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Fast green dye</td>
<td>SIGMA</td>
<td>F7252-5G&nbsp;&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Femtojet 4i Microinjector</td>
<td>Eppendorf</td>
<td>5252000013</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Glass capillaries of 1 mm diameter</td>
<td>WPI</td>
<td>TW100F-4</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Matrigel (extracellular matrix)</td>
<td>Corning</td>
<td>356237</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Microfuge tube 1.5ml</td>
<td>Eppendorf</td>
<td>T9661-1000EA</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Micro-loader tips</td>
<td>Eppendorf</td>
<td>612-7933</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Micropipette puller P-97</td>
<td>Shutter instrument</td>
<td>P-97</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Normal donkey Serum</td>
<td>Bio-Rad</td>
<td>C06SB</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Penstrep</td>
<td>Gibco</td>
<td>15140-122</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Sodium hypoclorite (use 5%)</td>
<td>Clorox</td>
<td>50371478</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Super stick slides</td>
<td>Waterborne, Inc</td>
<td>S100-3</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Swinnex-25 47mm Polycarbonate filter holder</td>
<td>EMD-Millipore</td>
<td>SX0002500</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Taurocholic acid sodium salt hydrate</td>
<td>SIGMA</td>
<td>T4009-5G</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Tween-20</td>
<td>Merck</td>
<td>8221840500</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Vectashield mounting agent</td>
<td>Vector Labs</td>
<td>H-1000</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Vortex Genie 2</td>
<td>Scientific industries, Inc</td>
<td>SI0236</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Adv+++ (DMEM+Penstrep+Glutamax+Hepes)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Final amount</td>
</tr>
<tr>
<td>DMEM</td>
<td>Invitrogen</td>
<td>12634-010</td>
<td>500ml</td>
</tr>
<tr>
<td>Penstrep</td>
<td>Gibco</td>
<td>15140-122</td>
<td>5ml of stock in 500ml DMEM</td>
</tr>
<tr>
<td>Glutamax</td>
<td>Gibco</td>
<td>35050038</td>
<td>5ml of stock in 500ml DMEM</td>
</tr>
<tr>
<td>Hepes</td>
<td>Gibco</td>
<td>15630056</td>
<td>5ml of stock in 500ml DMEM</td>
</tr>
<tr>
<td> INTESTINAL ORGANOID MEDIA-OME (Expansion media)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Final concentration</td>
</tr>
<tr>
<td>A83-01</td>
<td>Tocris</td>
<td>2939-50mg</td>
<td>0.5&micro;M</td>
</tr>
<tr>
<td>Adv+++</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>make upto 100 ml</td>
</tr>
<tr>
<td>B27</td>
<td>Invitrogen</td>
<td>17504044</td>
<td>1X</td>
</tr>
<tr>
<td>EGF</td>
<td>Peprotech</td>
<td>AF-100-15</td>
<td>50ng/mL</td>
</tr>
<tr>
<td>Gastrin</td>
<td>Tocris</td>
<td>3006-1mg</td>
<td>10 nM</td>
</tr>
<tr>
<td>NAC</td>
<td>Sigma</td>
<td>A9125-25G</td>
<td>1.25mM</td>
</tr>
<tr>
<td>NIC</td>
<td>Sigma</td>
<td>N0636-100G</td>
<td>10mM</td>
</tr>
<tr>
<td>Noggin CM</td>
<td>In house*</td>
<td>&nbsp;</td>
<td>10%</td>
</tr>
<tr>
<td>P38 inhibitor (SB202190)</td>
<td>Sigma</td>
<td>S7076-25 mg</td>
<td>10&micro;M</td>
</tr>
<tr>
<td>PGE2</td>
<td>Tocris</td>
<td>2296/10</td>
<td>10 nM</td>
</tr>
<tr>
<td>Primocin</td>
<td>InvivoGen</td>
<td>ant-pm-1</td>
<td>1ml/500ml media</td>
</tr>
<tr>
<td>RSpoI CM</td>
<td>In house*</td>
<td>&nbsp;</td>
<td>20%</td>
</tr>
<tr>
<td>Wnt3a CM</td>
<td>In house*</td>
<td>&nbsp;</td>
<td>50%</td>
</tr>
<tr>
<td>In house* - cell lines will be provided upon request</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>INTESTINAL ORGANOID MEDIA-OMD (Differentiation media)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>To differentiate organoids, expanding small intestinal organoids were grown in a Wnt-rich medium for six to seven days after splitting, and then grown in a differentiation medium (withdrawal of Wnt, nicotinamide, SB202190, in a differentiation medium (withdrawal of Wnt, nicotinamide, SB202190, prostaglandin E2 from a Wnt-rich medium or OME)</td>
</tr>
<tr>
<td>LUNG ORGANOID MEDIA- LOM (Differentiation media)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Final concentration</td>
</tr>
<tr>
<td>Adv+++</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>make upto 100 ml</td>
</tr>
<tr>
<td>ALK-I A83-01</td>
<td>Tocris</td>
<td>2939-50mg</td>
<td>500nM</td>
</tr>
<tr>
<td>B27</td>
<td>Invitrogen</td>
<td>17504044</td>
<td>0.0763888889</td>
</tr>
<tr>
<td>FGF-10</td>
<td>Peprotech</td>
<td>100-26</td>
<td>100ng/ml</td>
</tr>
<tr>
<td>FGF-7</td>
<td>Peprotech</td>
<td>100-19</td>
<td>25ng/ml</td>
</tr>
<tr>
<td>N-Acetylcysteine</td>
<td>Sigma</td>
<td>A9125-25G</td>
<td>1.25mM</td>
</tr>
<tr>
<td>Nicotinamide</td>
<td>Sigma</td>
<td>N0636-100G</td>
<td>5mM</td>
</tr>
<tr>
<td>Noggin UPE</td>
<td>U-Protein Express</td>
<td>Contact company directly</td>
<td>10%</td>
</tr>
<tr>
<td>p38 MAPK-I</td>
<td>Sigma</td>
<td>S7076-25 mg</td>
<td>1&micro;M</td>
</tr>
<tr>
<td>Primocin</td>
<td>InvivoGen</td>
<td>ant-pm-1</td>
<td>1:500</td>
</tr>
<tr>
<td>RhoKI Y-27632</td>
<td>Abmole Bioscience</td>
<td>M1817_100 mg</td>
<td>2.5&micro;m</td>
</tr>
<tr>
<td>Rspo UPE</td>
<td>U-Protein Express</td>
<td>Contact company directly</td>
<td>10%</td>
</tr>
<tr>
<td>Reducing buffer (for resuspension of oocysts and sporozoites for injection)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Final concentration</td>
</tr>
<tr>
<td>L-Glutathione reduced</td>
<td>Sigma</td>
<td>G4251-10MG</td>
<td>0.5 &mu;g/&mu;l of OME/OMD/LOM</td>
</tr>
<tr>
<td>Betaine</td>
<td>Sigma</td>
<td>61962</td>
<td>0.5 &mu;g/&mu;l of OME/OMD/LOM</td>
</tr>
<tr>
<td>L-Cysteine</td>
<td>Sigma</td>
<td>168149-2.5G</td>
<td>0.5 &mu;g/&mu;l of OME/OMD/LOM</td>
</tr>
<tr>
<td>Linoleic acid</td>
<td>Sigma</td>
<td>L1376-10MG</td>
<td>6.8 &mu;g/ml of OME/OMD /LOM</td>
</tr>
<tr>
<td>Taurine</td>
<td>Sigma</td>
<td>T0625-10MG</td>
<td>0.5 &mu;g/&mu;l of OME/OMD/LOM</td>
</tr>
<tr>
<td>Blocking buffer (for immunoflourescence staining)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Final concentration</td>
</tr>
<tr>
<td>Donkey/Goat serum</td>
<td>Bio-Rad</td>
<td>C06SB</td>
<td>2%</td>
</tr>
<tr>
<td>PBS</td>
<td>Thermo-Fisher</td>
<td>70011044</td>
<td>Make upto 100ml</td>
</tr>
<tr>
<td>Tween 20</td>
<td>Merck</td>
<td>P1379</td>
<td>0.1%</td>
</tr>
<tr>
<td>List of Antibodies used </td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
</tr>
<tr>
<td>Alexa 568 goat anti-rabbit</td>
<td>Invitrogen</td>
<td>A-11011</td>
<td>Dilution-1:500; RRID: AB_143157</td>
</tr>
<tr>
<td>Crypt-a-Glo Comprehensive Kit- Fluorescein-labeled antibody Crypto-Glo</td>
<td>Waterborne, Inc</td>
<td>A400FLK</td>
<td>Dilution- 1:200</td>
</tr>
<tr>
<td>Crypta-Grab IMS Beads- Magnetic beads coated in monoclonal antibody reactive</td>
<td>Waterborne, Inc</td>
<td>IMS400-20</td>
<td>Dilution-1:500</td>
</tr>
<tr>
<td>DAPI</td>
<td>Thermo Fisher Scientific</td>
<td>D1306</td>
<td>Dilution-1:1000; RRID : AB_2629482</td>
</tr>
<tr>
<td>Phalloidin-Alexa 674</td>
<td>Invitrogen</td>
<td>A22287</td>
<td>Dilution-1:1000; RRID: AB_2620155</td>
</tr>
<tr>
<td>Rabbit anti-gp15 antibody generated by R. M. O'Connor (co-author).</td>
<td>Upon request</td>
<td>Upon request</td>
<td>Dilution-1:500</td>
</tr>
<tr>
<td>Sporo-Glo</td>
<td>Waterborne, Inc</td>
<td>A600FLR-1X</td>
<td>Dilution- 1:200</td>
</tr>
</tbody>
</table>

studying cryptosporidium infection in 3d tissue-derived human organoid culture systems by microinjection

隐孢子虫是导致人类腹泻病的主要原因之一。为了了解寄生虫的病理学并开发有效的药物，需要一种体外培养系统来概括宿主的条件。有机体与它们起源的组织非常相似，是研究宿主-寄生虫相互作用的理想。有机体是三维（3D）组织衍生结构，从成人干细胞中提取，在培养中长期生长，无需任何基因畸变或转化。它们具有明确的极性，具有尖顶和边面。有机体在药物测试、生物库、疾病建模和宿主微生物相互作用研究方面具有多种应用。在这里，我们提出了一个分步协议，如何准备隐孢子虫的卵母虫和孢子菌感染人类肠道和气道器官。然后，我们演示了如何使用微注射将微生物注射到有机体流明中。有机物可用于宿主-微生物相互作用研究（显微注射、机械剪切和电镀以及制造单层）有三种主要方法。微注射能够维护3D结构，并能够精确控制微生物的寄生虫体积和直接的侧接触。我们提供用于成像或卵母细胞生产的最佳器官生长的详细信息。最后，我们还演示了如何从有机体中分离出新生成的卵母细胞，以便进行进一步的下游处理和分析。

这里介绍的协议导致卵囊和孢子石的有效纯化（图1A）准备显微注射。排泄物协议导致从大约 70-80% 的卵母细胞中释放孢子菌，因此必须通过 3 μm 过滤器过滤掉剩余的卵母细胞和壳。过滤可产生近100%的孢子石纯化（图1B）。此外，添加绿色染料有助于确保注射所有有机物，并允许注射的有机物在注射后至少24小时可视化（图2B）。<...

Watch this Scientific Journal Video about Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection at JoVE.com

Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection

我们描述了用于制备卵母细胞和纯化孢子菌的方案，用于研究隐孢子虫的人类肠道和气道器官的感染。我们演示了将寄生虫微注射到肠道器官流明和有机体免疫染色的程序。最后，我们描述了从器官中分离生成的卵母细胞。

通过微注射研究3D组织衍生的人体器官培养系统中的隐孢子虫感染

Cryptosporidium parvum is one of the major causes of human diarrheal disease. To understand the pathology of the parasite and develop efficient drugs, an ...

使用这种孢子体纯化和微注射程序，基本上任何宿主微生物都可以在任何组织衍生器官系统中进行研究。使用这种技术，我们可以控制注入器官流明或器官的一面的纯孢子体的数量。要纯化C.parvum孢子体，将卵母细胞转移到15毫升管中，然后以每毫升外站介质浓度的1倍10再给第七个卵母细胞。在37摄氏度下60至90分钟后，将14毫升适当的洗涤溶液与卵母细胞混合，通过离心沉淀细胞。小心吸制上流液，避免失去卵母细胞，并在每一到两毫升DMEM浓度的三倍10至第七粒中重新暂停孢子球。要去除剩余的卵母体和壳体，请将装有 47 毫升滤芯支架装置的 10 毫升注射器筒放入 4 摄氏度的 15 毫升管中。向过滤器组件中加入 7.5 毫升的孢子石悬浮液。当整个体积通过过滤器的重力，洗净任何剩余的细胞通过过滤器与另外7.5毫升的DMEM。当所有洗涤液都经过过滤器时，通过离心收集过滤的孢子石悬浮液10分钟，并在50至100微升适当的有机体培养基中标记颗粒，辅以0.05%的快速绿色染料，以及L-谷胱甘肽、贝塔因、L-半胱氨酸、亚油酸和含有还原缓冲的牛磺酸。要将寄生虫微注射到 3D 器官的一侧，首先使用微移子拉拔器准备玻璃注射毛细管。使用钳子将毛细管的尖端切割到 9 到 12 微米直径，以便于流水杨石或卵石，如果您选择直接注射卵母质，并使用微装载机尖端填充每个毛细管，并填充标有卵石或孢子石悬浮液的快速绿色染料。然后，将第一个充满孢子的毛细管装入显微显胶中，并使用倒置显微镜以 5 倍的放大倍率和恒定的压力将悬浮液的 100 到 200 纳米光插入每个器官中。外站协议导致从大约 70% 到 80% 的卵石释放孢子虫。因此，通过三微米过滤过滤掉剩余的卵壳至关重要。去除近100%的未切除的卵母细胞。此外，添加绿色染料有助于确保注射所有器官，并允许在注射后至少24小时内对注射的器官进行可视化。虽然这是一种行之有效的方法，但扫描电子显微镜可用于确保外显器过程不会损坏孢子虫或卵石。向器官流明注射等量的卵母线可以通过简单的显微成像得到视觉确认。免疫荧光分析也可用于探索受隐孢子虫感染的细胞类型。分化的有机体被感染五天后，通过将部分卵母细胞干燥到粘合剂幻灯片上，用甲醇固定卵母细胞，将DAPI染色与合适的卵母细胞特异性抗体相结合，可以确认卵母细胞内存在孢子体的存在。为了获得最佳效果，请使用新鲜的卵巢，使用相同的毛细管进行所有注射，并在注射后每天更换介质。使用这种技术，各种实验室现在开始使用器官来研究宿主-微生物的相互作用。我们正在研究类似的有机体微培养环境中的共体和其他病原体。由于隐孢子虫是一种人类寄生虫，因此按照2级安全规定进行这些实验非常重要。

Research

JoVE Journal

Immunology and Infection

Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection

Recurrent Herpetic Stromal Keratitis in Mice, a Model for Studying Human HSK

复发性单纯疱疹性角膜基质性角膜炎小鼠模型研究人类HSK

Herpetic eye disease, termed herpetic stromal keratitis (HSK), is a potentially blinding infection of the cornea that results in over 300,000 clinical ...

科研

免疫与感染

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.

评估的HIV-1感染的CD4的先天传感<sup&gt; +</sup&gt;性T细胞浆细胞样树突细胞使用<em&gt;体外</em&gt;共培养体系。

HIV-1 innate sensing requires direct contact of infected CD4+ T cells with plasmacytoid dendritic cells (pDCs). In order to study this process, the ...

Organoids as Model for Infectious Diseases: Culture of Human and Murine Stomach Organoids and Microinjection of Helicobacter Pylori

类器官的模型传染病：人类的文化与小鼠胃组织体与幽门螺杆菌的显微注射

Recently infection biologists have employed stem cell derived cultures to answer the need for new and better models to study host-pathogen interactions. ...

Mouse- and Human-derived Primary Gastric Epithelial Monolayer Culture for the Study of Regeneration

小鼠和人源性胃上皮单层培养对再生的研究

In vitro studies of gastric wound repair typically involves the use of gastric cancer cell lines in a scratch-wound assay of cellular proliferation and ...

Quantification of Efferocytosis by Single-cell Fluorescence Microscopy

单细胞荧光显微镜定量 Efferocytosis 的研究

Studying the regulation of efferocytosis requires methods that are able to accurately quantify the uptake of apoptotic cells and to probe the signaling ...

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction

实时定量聚合酶链反应检测感染细胞培养中的登革热病毒 rna

At present, real-time polymerase chain reaction (PCR) technology is an indispensable tool for the detection and quantification of viral genomes in ...

Analysis of Interactions between Endobiotics and Human Gut Microbiota Using In Vitro Bath Fermentation Systems

内生菌与人类肠道微生物群使用体外浴发酵系统的相互作用分析

Human intestinal microorganisms have recently become an important target of research in promoting human health and preventing diseases. Consequently, ...

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

高产细胞外囊泡制剂的纯化远离病毒

One of the major hurdles in the field of extracellular vesicle (EV) research today is the ability to achieve purified EV preparations in a viral infection ...

Pneumococcus Infection of Primary Human Endothelial Cells in Constant Flow

恒流中原发性人类内皮细胞的肺炎球菌感染

Interaction of Streptococcus pneumoniae with the surface of endothelial cells is mediated in blood flow via mechanosensitive proteins such as the Von ...

Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry

Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry

利用流细胞测定人类白细胞测定阿斯珀吉鲁斯紫锥体烟曲的噬菌体

Invasive pulmonary infection by the mold Aspergillus fumigatus poses a great threat to immunocompromised patients. Inhaled fungal conidia (spores) are ...