Name: an in vitro model of a parallel-plate perfusion system to study bacterial adherence to graft tissues
Uploaded: 2019-01-07T14:00:00
Description: Watch this Scientific Journal Video about An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues at JoVE.com

这项研究是由向生殖健康提供的研究基金 ku leuven (ot/14/197) 的赠款赞助的。trv 是佛兰德研究基金会的博士后研究员 (比利时;grant 编号-12k0916n) 和 rh 由 uz leuven 的临床研究基金提供支助。

1. 为体外研究准备移植组织
注: 使用了三种类型的组织: 牛心包贴片 (bp)、冷冻保留同种异体移植 (ch) 和牛颈静脉移植 (bjv)。在 bjv 导管和 ch (由欧洲同种异体移植银行 (ehb) 处理并在使用前储存在液氮中的组织) 的情况下, 使用壁面和瓣膜小叶。从制造商处购买了 bp 贴片和 bjv 管道。使用前, 按照 ehb 指令12 解冻 ch。
<ol><li>使用前使用0.9% 的氯化钠冲洗所...

<ol>
	<li>Que, Y. A., Moreillon, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infective+endocarditis.">Infective endocarditis.</a> Nature Reviews Cardiology. 8 (6), 322-336 (2011).</li><li>Werdan, K., 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=Mechanisms+of+infective+endocarditis:+pathogen-host+interaction+and+risk+states.">Mechanisms of infective endocarditis: pathogen-host interaction and risk states.</a> Nature Reviews Cardiology. 11 (1), 35-50 (2014).</li><li>Moreillon, P., Que, Y. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infective+endocarditis.">Infective endocarditis.</a> The Lancet. 363 (9403), 139-149 (2004).</li><li>Jalal, Z., 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=Selective+propensity+of+bovine+jugular+vein+material+to+bacterial+adhesions:+An+in+vitro+study.">Selective propensity of bovine jugular vein material to bacterial adhesions: An in vitro study.</a> International Journal of Cardiology. 198, 201-205 (2015).</li><li>Sharma, A., Cote, A. T., Hosking, M. C. K., Harris, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Systematic+Review+of+Infective+Endocarditis+in+Patients+With+Bovine+Jugular+Vein+Valves+Compared+With+Other+Valve+Types.">A Systematic Review of Infective Endocarditis in Patients With Bovine Jugular Vein Valves Compared With Other Valve Types.</a> JACC Cardiovascular Interventions. 10 (14), 1449-1458 (2017).</li><li>Malekzadeh-Milani, S., 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=Incidence+and+predictors+of+Melody(R)+valve+endocarditis:+a+prospective+study.">Incidence and predictors of Melody(R) valve endocarditis: a prospective study.</a> Archives of Cardiovascular Diseases. 108 (2), 97-106 (2015).</li><li>Hill, E. E., 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=Management+of+prosthetic+valve+infective+endocarditis.">Management of prosthetic valve infective endocarditis.</a> American Journal of Cardiology. 101 (8), 1174-1178 (2008).</li><li>Claes, J., 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=Clumping+factor+A,+von+Willebrand+factor-binding+protein+and+von+Willebrand+factor+anchor+Staphylococcus+aureus+to+the+vessel+wall.">Clumping factor A, von Willebrand factor-binding protein and von Willebrand factor anchor Staphylococcus aureus to the vessel wall.</a> Journal of Thrombosis and Haemostasis. 15 (5), 1009-1019 (2017).</li><li>Fowler, T., 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=Cellular+invasion+by+Staphylococcus+aureus+involves+a+fibronectin+bridge+between+the+bacterial+fibronectin-binding+MSCRAMMs+and+host+cell+beta1+integrins.">Cellular invasion by Staphylococcus aureus involves a fibronectin bridge between the bacterial fibronectin-binding MSCRAMMs and host cell beta1 integrins.</a> European Journal of Cell Biology. 79 (10), 672-679 (2000).</li><li>Patti, J. M., Hook, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbial+adhesins+recognizing+extracellular+matrix+macromolecules.">Microbial adhesins recognizing extracellular matrix macromolecules.</a> Current Opinion in Cell Biology. 6 (5), 752-758 (1994).</li><li>Massey, R. 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=Fibronectin-binding+protein+A+of+Staphylococcus+aureus+has+multiple,+substituting,+binding+regions+that+mediate+adherence+to+fibronectin+and+invasion+of+endothelial+cells.">Fibronectin-binding protein A of Staphylococcus aureus has multiple, substituting, binding regions that mediate adherence to fibronectin and invasion of endothelial cells.</a> Cellular Microbiology. 3 (12), 839-851 (2001).</li><li>Jashari, 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=Belgian+and+European+experience+with+the+European+Homograft+Bank+(EHB)+cryopreserved+allograft+valves--assessment+of+a+20+year+activity.">Belgian and European experience with the European Homograft Bank (EHB) cryopreserved allograft valves--assessment of a 20 year activity.</a> Acta Chirurgica Belgica. 110 (3), 280-290 (2010).</li><li>Cheatham, J. 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=Clinical+and+hemodynamic+outcomes+up+to+7+years+after+transcatheter+pulmonary+valve+replacement+in+the+US+melody+valve+investigational+device+exemption+trial.">Clinical and hemodynamic outcomes up to 7 years after transcatheter pulmonary valve replacement in the US melody valve investigational device exemption trial.</a> Circulation. 131 (22), 1960-1970 (2015).</li><li>Que, Y. 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=Fibrinogen+and+fibronectin+binding+cooperate+for+valve+infection+and+invasion+in+Staphylococcus+aureus+experimental+endocarditis.">Fibrinogen and fibronectin binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis.</a> The Journal of Experimental Medicine. 201 (10), 1627-1635 (2005).</li><li>Veloso, T. 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=Bacterial+adherence+to+graft+tissues+in+static+and+flow+conditions.">Bacterial adherence to graft tissues in static and flow conditions.</a> The Journal of Thoracic and Cardiovascular Surgery. 155 (1), 325-332 (2018).</li><li>Liesenborghs, L., Verhamme, P., Vanassche, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Staphylococcus+aureus,+master+manipulator+of+the+human+hemostatic+system.">Staphylococcus aureus, master manipulator of the human hemostatic system.</a> Journal of Thrombosis and Haemostasis. 16 (3), 441-454 (2018).</li><li>Chiu, J. J., 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=Shear+stress+increases+ICAM-1+and+decreases+VCAM-1+and+E-selectin+expressions+induced+by+tumor+necrosis+factor-[alpha]+in+endothelial+cells.">Shear stress increases ICAM-1 and decreases VCAM-1 and E-selectin expressions induced by tumor necrosis factor-[alpha] in endothelial cells.</a> Artheriosclerosis, Thrombosis, and Vascular Biology. 24 (1), 73-79 (2004).</li><li>Jockenhoevel, S., Zund, G., Hoerstrup, S. P., Schnell, A., Turina, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiovascular+tissue+engineering:+a+new+laminar+flow+chamber+for+in+vitro+improvement+of+mechanical+tissue+properties.">Cardiovascular tissue engineering: a new laminar flow chamber for in vitro improvement of mechanical tissue properties.</a> ASAIO Journal. 48 (1), 8-11 (2002).</li><li>Veltrop, M. H. A. 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=Bacterial+Species-+and+Strain-Dependent+Induction+of+Tissue+Factor+in+Human+Vascular+Endothelial+Cells.">Bacterial Species- and Strain-Dependent Induction of Tissue Factor in Human Vascular Endothelial Cells.</a> Infection and Immunity. 67 (11), 6130-6138 (1999).</li></ol>

最近的临床观察使 ie 作为一种并发症的患者接受瓣膜置换术 rvot6,13。ie 中植入瓣膜功能障碍是细菌与血管内移植物相互作用的结果, 导致广泛的炎症和凝血反应1,14。提出的新的体外模型使我们能够研究组织结构和细菌因素的差异是否可能调节体内使用的移植物15对感染的易感性.</sup...

金黄色葡萄球菌(金黄色葡萄球菌) 采用了各种毒力策略来规避宿主免疫防御系统, 使植入人体循环的生物或非生物表面殖民, 从而导致严重的血管内感染, 如败血症和ie1,2,3,4,5。ie 仍然是一个重要的治疗相关并发症的患者植入假体心脏瓣膜后, 而个别因素的原因, 导致 ie7 的发病还没有完全了解 6,7。在流动条件下, 细菌会遇到剪切力, 为了粘附在容器壁8 上, 它们需要克服剪切力。模型可以研究细菌和假体瓣膜组织或内皮在流动中的相互作用, 因为它们更多地反映了体内的情况。
几种特殊的机制促进细菌粘附到内皮细胞 (ecs) 和暴露的内皮细胞下基质 (ecm), 导致组织定植和植物成熟, 是 ie9中必不可少的早期步骤。各种葡萄球菌表面蛋白或 mscram (识别粘附基质分子的微生物表面成分) 被描述为通过与纤维连接蛋白等分子相互作用而与宿主细胞和 ecm 蛋白粘附的介质,纤维蛋白原、胶原蛋白和冯·威利布兰德因子 (vwf)8、10、11。然而, 鉴于分子内折叠的一些毒力因素, 大多在静态条件下研究, 其中许多相互作用可能有不同的相关性, 在循环血液的血管内感染。
因此, 我们提出了一个内部设计的体外平行板流动室模型, 允许评估细菌在组织移植中植入 rvot 位置时对 ecm 和 ecs 不同成分的粘附情况。本研究中描述的方法的总体目的是研究细菌与潜在血管内组织在流动条件下的相互作用机制, 这些机制与血液病原体的体内环境密切相关, 如s. 金牛座.这种新的方法侧重于移植组织表面对细菌粘附的敏感性, 以确定影响 ie 发展的潜在危险因素。

<table>
	<tbody>
		<tr>
			<td>Bovine Pericardium (BP) patch, Supple Peri-Guard Pericardium</td>
			<td>Synovis Surgical Innovations, USA</td>
			<td>PC-0404SN</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Jugular Vein conduits (BJV)</td>
			<td>Contegra conduit; Medtronic Inc, USA</td>
			<td>M333105D001</td>
			<td></td>
		</tr>
		<tr>
			<td>CH cryopreserved homograft</td>
			<td>European Homograft Bank (EHB)</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Acu-Punch</td>
			<td>Acuderm Inc, USA</td>
			<td>P850 (8 mm); P1050 (10 mm)</td>
			<td></td>
		</tr>
		<tr>
			<td>human Albumin</td>
			<td>Flexbumin; Baxter, Belgium</td>
			<td>BE171464 
			LOT:16G12C</td>
			<td></td>
		</tr>
		<tr>
			<td>Tryptic soy broth (TSB)</td>
			<td>Fluka, Steinheim, Germany</td>
			<td>22092-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Heart infusion broth (BHI)</td>
			<td>Fluka</td>
			<td>53286-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffered saline (PBS).</td>
			<td>Gibco</td>
			<td>14190-094</td>
			<td></td>
		</tr>
		<tr>
			<td>5(6)-Carboxyfluorescein N-hydroxysuccinimide ester (CF)</td>
			<td>Sigma-Aldrich, Germany</td>
			<td>21878-100MG-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Peristaltic pump (MODEL ISM444B)</td>
			<td>Ismatec BVP-Z Standard; Cole Parmer, Wertheim, Germany</td>
			<td>631942-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Sonication bath</td>
			<td>VWR Ultrasonic Cleaner; VWR, Radnor, Pa</td>
			<td>142-6044</td>
			<td>230V/50 -60Hz 60VA; HF45kHz, 30W</td>
		</tr>
		<tr>
			<td>ProLong Gold Antifade Mountant</td>
			<td>Invitrogen by ThermoFisher</td>
			<td>P36930</td>
			<td></td>
		</tr>
		<tr>
			<td>InCell Analyzer 2000 (fluorescence scanner)</td>
			<td>GE Healthcare Life Sciences, Pittsburgh, Pa</td>
			<td>29027886</td>
			<td></td>
		</tr>
		<tr>
			<td>Arium Pro VF - ultrapure water - H2O MilliQ</td>
			<td>Millipore</td>
			<td>87206462</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscopic slides - Tissue Culture Chambers (1-well)</td>
			<td>Sarstedt</td>
			<td>94.6140.102</td>
			<td></td>
		</tr>
		<tr>
			<td>1-well on Lumox detachable</td>
			<td>Sarstedt</td>
			<td>94.6150.101</td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless Steel - surgical Blades</td>
			<td>Swann-Morton</td>
			<td>311</td>
			<td></td>
		</tr>
		<tr>
			<td>Tygon Silicone Tubing, 1/8&#34;ID x 1/4&#34;OD</td>
			<td>Cole-Parmer</td>
			<td>EW-95702-06</td>
			<td>Temperature range: &#8211;80 to 200&#176;C 
			Sterilize: With ethylene oxide, gamma irradiation, or autoclave for 30 min, 15 psi of pressure</td>
		</tr>
		<tr>
			<td>PharMed BPT Tubing</td>
			<td>Saint-Gobain</td>
			<td>AY242012</td>
			<td>Autoclavable 30 min at 121&#176;C</td>
		</tr>
		<tr>
			<td>Tygon LMT-55 Tubing</td>
			<td>Saint Gobain Performance Plastics&#8482;</td>
			<td>15312022</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermostat</td>
			<td>BMG BIOMEDIZINTECHNIK</td>
			<td>300-0042</td>
			<td>230V, 90VA, 50Hz</td>
		</tr>
	</tbody>
</table>

an in vitro model of a parallel-plate perfusion system to study bacterial adherence to graft tissues

各种瓣膜导管和支架安装的瓣膜用于先天性心脏病患者的右心室流出道 (rvot) 瓣膜置换术。然而, 当使用假肢材料时, 这些移植物容易受到细菌感染和各种宿主反应。
识别在微生物血管内粘连中发挥重要作用的细菌和宿主因素对于更好地了解感染性心内膜炎 (ie) 等感染的病理生理学和开展预防工作具有重要意义策略。因此, 有必要建立有效的模型来研究细菌在生理剪切条件下的粘附情况。在这里, 我们描述了一个新设计的基于平行板的体外灌注室的使用情况, 该室允许研究细菌对不同器官移植组织的粘附性, 如暴露的细胞外基质、内皮细胞和惰性区域.该方法结合成菌区量值, 可用于评价移植物材料在流动条件下对细菌粘附的倾向。此外, 流室系统还可用于研究血液成分在剪切条件下细菌粘附中的作用。我们证明, 组织的来源, 其表面形态和细菌物种特异性不是主要的决定细菌坚持到移植组织的因素, 使用我们的内部设计的体外灌注模型。

为了更好地了解 ie 发育背后的机制, 该模型能够评估感染发病体内存在的细菌和组织相关因素。
在详细的, 新的体外方法允许量化细菌在流动条件下对不同的移植组织的粘附, 在施加剪切应力的组织上灌注荧光标记的细菌, 在3-10 的生理范围内施加剪切应力/cm2为 rvot。在这项工作中, 我们使用了 4 mL/mi...

Watch this Scientific Journal Video about An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues at JoVE.com

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

我们描述了一个内部设计的体外流室模型, 它允许研究细菌对移植组织的粘附。

平行板灌注系统的体外模型研究了细菌对移植组织的粘附性

Various valved conduits and stent-mounted valves are used for right ventricular outflow tract (RVOT) valve replacement in patients with congenital heart ...

这种方法可以帮助回答感染性心内膜炎的发病机制中的关键问题，以及通过细菌、血细胞和内皮细胞内衬肌肉等领域进行病理图谱。该技术的主要优点是，组织移植物可能直接暴露在与各种靶点（如细菌、血小板和蛋白质）的相互作用中，在标准化的流条件下。这项技术的含意延伸到我们的感染性内膜炎的治疗，因为它可以解开重要的分子相互作用和因素，推动某些组织的高感染性。该方法提供了植被形成的发病机制的见解。它也可用于研究血管渗透性、细胞流动性和基因细胞表达。为了开始该协议，放置以前准备的组织活检直径10毫米，显微镜幻灯片和8毫米圆形穿孔之间的相同厚度，以及内表面朝上处的橡胶垫片，以接触细菌悬浮液。在支架中放置组织移植方向时，重点关注。确保组织的内部表面已经暴露在血液表面向上接触您的实验介质。接下来，将带组织的支架插入嵌入在腔室底部金属框架中的垫片中。将带相应垫片的上部金属框架与先前插入的组织支架一起连接到腔室底部。然后用八个螺钉和螺钉安装整个腔室。使用卡钳或尺子确保腔室高度在嫁接时始终相同。接下来，用近体泵连接流量室。然后将 400 毫升储液罐与管连接。将100毫升悬浮液（10升）吸向磷酸盐缓冲盐水中的7个荧光标记菌落形成单元的组织，其绝对应力为每平方厘米3餐，使用心肺泵和细菌储液库，每分钟4毫升的相应流量为1小时。使用同一收集储液罐连续循环 100 毫升细菌悬浮液。灌注后，拆除腔室以释放移植物。使用轨道摇床在12井板中用4毫升PBS清洗组织两次，进行三分钟。然后使用直径较小的皮肤活检冲孔切割移植物的内部部分。将每个组织活检放入一个单独的14毫升管中，其中含有一毫升无菌0.9%氯化钠。使用声波浴将细菌从组织中分离出来10分钟。准备一个14毫升管与10毫升无菌0.9%氯化钠，使连续稀释后获得的细菌悬浮液声波。标记管号 2。准备三个14毫升管与10毫升无菌0.9%氯化钠，用于连续稀释的初始细菌悬浮液到灌注实验。标记管号 3、4 号和 5 号。接下来，固定三个加糖板，一个用于细菌灌注，一个用于控制PBS灌注，第三个用于最初用于灌注的细菌悬浮液。以下列方式标记每个板的三个扇区：10-1、10-3 和 10-4。要计算细菌中形成菌落的单位数，请将板标为：10-1、10-3、10-5 和 10-7。旋转含有组织活检的管子，每个15秒，使连续稀释。要准备串行稀释，将100微升管1号转移到管号二号，并涡旋管彻底混合溶液。将100微升的管号和二号管的内容物扩散到相应的扇区，10-1和10-3的加糖板。然后将10号管的10微升在10号四区。重复此步骤四次，从每卷 10 微升中获取四个单独的增长。为了准备初始培养物的连续稀释，首先涡流稀释15秒，将100微升细菌悬浮液转移到三号管，然后通过涡流进行大力混合。将 100 微升的三号管添加到管四中，然后再次混合。从管号四重复管号 5 的过程。将三号、四号、五号管和标记的初始培养物的100微升内容分别扩散到血脂板的10-3、10-5、10-7和10-1的部门。将血脂板放在层罩下，以空气干燥细菌扩散，通常持续 10 分钟。之后，将板在37摄氏度下放置，进行过夜孵育。经过一夜的孵育，计算细菌菌落，以获得CFO的数量。在纯粹的压力条件下，观察到金黄色葡萄球菌和金黄色葡萄球菌表皮菌感染在各种移植组织之间出现类似的细菌附着。按照程序，与牛的葡萄球菌斑块相比，桑吉诺丝对牛的血管壁的依从性明显减少。在比较这三种细菌时，桑吉诺氏链球菌与金黄色葡萄球菌和金黄色葡萄球菌表皮菌相比，与牛皮脉壁的粘附性显著降低。在尝试此程序时，重要的是要记住要准备同一头部的正确组织。它将确保所有实验样本的类似条件，并最大限度地减少不同系列实验之间的数据变异性。开发后，该技术允许研究人员通过丰富其体外模型方法，以不同的元素建立接近医疗状态的全部复杂性，从而逐步探索复杂的疾病系统。

Research

JoVE Journal

Developmental Biology

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

A Novel Ex Ovo Banding Technique to Alter Intracardiac Hemodynamics in an Embryonic Chicken System

A Novel Ex Ovo Banding Technique to Alter Intracardiac Hemodynamics in an Embryonic Chicken System

一本小说<em&gt;防爆大毛</em&gt;带技术来改变心内血流动力学在鸡胚胎系统

The new model presented here can be used to understand the influence of hemodynamics on specific cardiac developmental processes, at the cellular and ...

科研

发育生物学

Development of an Ethanol-induced Fibrotic Liver Model in Zebrafish to Study Progenitor Cell-mediated Hepatocyte Regeneration

乙醇引起的肝纤维化模型的斑马鱼发展研究祖细胞介导的​​肝细胞的再生

Sustained liver fibrosis with continuation of extracellular matrix (ECM) protein build-up results in the loss of cellular competency of the liver, leading ...

In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition

In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition

在体外和体内模型来研究角膜内皮-间充质转换

Corneal endothelial cells (CECs) play a crucial role in maintaining corneal clarity through active pumping. A reduced CEC count may lead to corneal edema ...

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 ...

Whole-mount Confocal Microscopy for Adult Ear Skin: A Model System to Study Neuro-vascular Branching Morphogenesis and Immune Cell Distribution

成人耳皮肤全安装共焦显微术: 一种研究神经血管分支形态发生和免疫细胞分布的模型系统

Here, we present a protocol of a whole-mount adult ear skin imaging technique to study comprehensive three-dimensional neuro-vascular branching ...

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 ...

A Hyperandrogenic Mouse Model to Study Polycystic Ovary Syndrome

Hyperandrogenic 小鼠模型研究多囊卵巢综合征

Hyperandrogenemia plays a critical role in reproductive and metabolic function in females and is the hallmark of polycystic ovary syndrome. Developing a ...

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 ...

Clonal Genetic Tracing using the Confetti Mouse to Study Mineralized Tissues

使用纸屑小鼠研究矿化组织的克隆遗传追踪

Labeling an individual cell in the body to monitor which cell types it can give rise to and track its migration through the organism or determine its ...

Whole-Mount In Situ Hybridization in Zebrafish Embryos and Tube Formation Assay in iPSC-ECs to Study the Role of Endoglin in Vascular Development

斑马鱼胚胎全山原位杂交和管状测定在iPSC-ECs中研究恩多林在血管发育中的作用

Vascular development is determined by the sequential expression of specific genes, which can be studied by performing in situ hybridization assays in ...