Name: extraction, labeling, and purification of lineage-specific cells from human antral follicles
Uploaded: 2022-11-30T13:40:00
Description: Watch this Scientific Journal Video about Extraction, Labeling, and Purification of Lineage-Specific Cells from Human Antral Follicles at JoVE.com

作者感谢Queenie Victorina Neri研究学者奖（D.J.）和刘洪清研究学者奖（L.M.）的支持。N.L.G得到了NYSTEM干细胞和再生医学博士后培训基金的支持。

所有涉及小鼠的程序均由威尔康奈尔医学院的机构动物护理和使用委员会（IACUC）批准。所有使用卵巢组织的异种移植实验均按照相关指南和规定进行。两个卵巢都是从一名14岁的脑死亡器官捐献者中分离出来的，该捐献者没有放疗/化疗史，也没有内分泌或生殖疾病记录。威尔康奈尔医学院的机构审查委员会（IRB）委员会批准了组织的采集，并在知情同意使用组织后获得了器官捐献者家属的批准?...

<ol>
	<li>Aghadavod, 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=Isolation+of+granulosa+cells+from+follicular+fluid;+Applications+in+biomedical+and+molecular+biology+experiments.">Isolation of granulosa cells from follicular fluid; Applications in biomedical and molecular biology experiments.</a> Advanced Biomedical Research. 4, 250 (2015).</li><li>Man, 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=Comparison+of+human+antral+follicles+of+xenograft+versus+ovarian+origin+reveals+disparate+molecular+signatures.">Comparison of human antral follicles of xenograft versus ovarian origin reveals disparate molecular signatures.</a> Cell Reports. 32 (6), 108027 (2020).</li><li>Schmidt, K. L., Ernst, E., Byskov, A. G., Nyboe Andersen, A., Yding Andersen, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival+of+primordial+follicles+following+prolonged+transportation+of+ovarian+tissue+prior+to+cryopreservation.">Survival of primordial follicles following prolonged transportation of ovarian tissue prior to cryopreservation.</a> Human Reproduction. 18 (12), 2654-2659 (2003).</li><li>Jensen, A. 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=Outcomes+of+transplantations+of+cryopreserved+ovarian+tissue+to+41+women+in+Denmark.">Outcomes of transplantations of cryopreserved ovarian tissue to 41 women in Denmark.</a> Human Reproduction. 30 (12), 2838-2845 (2015).</li><li>Newton, H., Aubard, Y., Rutherford, A., Sharma, V., Gosden, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low+temperature+storage+and+grafting+of+human+ovarian+tissue.">Low temperature storage and grafting of human ovarian tissue.</a> Human Reproduction. 11 (7), 1487-1491 (1996).</li><li>Man, 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=Chronic+superphysiologic+AMH+promotes+premature+luteinization+of+antral+follicles+in+human+ovarian+xenografts.">Chronic superphysiologic AMH promotes premature luteinization of antral follicles in human ovarian xenografts.</a> Science Advances. 8 (9), 7315 (2022).</li><li>Gougeon, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+follicular+growth+in+the+human:+A+model+from+preliminary+results.">Dynamics of follicular growth in the human: A model from preliminary results.</a> Human Reproduction. 1 (2), 81-87 (1986).</li><li>Richards, J. S., Ren, Y. A., Candelaria, N., Adams, J. E., Rajkovic, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+follicular+theca+cell+recruitment,+differentiation,+and+impact+on+fertility:+2017+update.">Ovarian follicular theca cell recruitment, differentiation, and impact on fertility: 2017 update.</a> Endocr Rev. 39 (1), 1-20 (2018).</li><li>Asiabi, P., Dolmans, M. M., Ambroise, J., Camboni, A., Amorim, C. A. <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+differentiation+of+theca+cells+from+ovarian+cells+isolated+from+postmenopausal+women.">In vitro differentiation of theca cells from ovarian cells isolated from postmenopausal women.</a> Human Reproduction. 35 (12), 2793-2807 (2020).</li><li>Dalman, A., Totonchi, M., Valojerdi, M. R. <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+human+theca+stem+cells+and+their+differentiation+into+theca+progenitor+cells.">Establishment and characterization of human theca stem cells and their differentiation into theca progenitor cells.</a> Journal of Cellular Biochemistry. 119 (12), 9853-9865 (2018).</li></ol>

由于几个原因，更好地解析卵巢卵泡内的细胞多样性在临床上很重要。在将上述方案应用于分离位于窦期卵泡内的独特表型亚型时，应考虑几个因素。首先，来源于窦卵泡的卵巢组织的健康和活力对于确定细胞的质量和下游应用的成功至关重要。这可以通过最小化缺血间隔、快速工作（最好成对）以及准备将用于组织处理和 FACS 的实验室空间来优化。其次，这些方案非常适合窦期卵泡，因为它们?...

人类卵巢的主要功能元素是卵泡，卵泡控制着卵母细胞的生长和发育。在体外受精的背景下，已经建立了分离滤泡细胞的方案，但这些方案仅适用于在卵母细胞提取时从黄 体 化卵泡中收集细胞1。我们开发了一种方案，可以在天然卵巢或异种移植卵巢组织产生的不同发育阶段从窦卵泡中分离离散细胞群2。尽管人们一致认为卵泡驻留细胞对卵母细胞培养的贡献非常重要，但很少有研究前瞻性地鉴定和提取窦期卵泡中存在的独特表型亚型。更深入地了解不同发育阶段特化细胞之间的分化层次结构和信号转导可以拓宽我们对稳态和病理条件下卵巢生理学的理解。此外，对离散细胞亚型的区分及其对卵泡生长/成熟的分子贡献可能提供一种产生离 体 替代物的方法，这些替代物重建卵巢功能以促进卵母细胞成熟和/或治疗内分泌功能障碍。 
卵巢内的每种独特细胞类型都有助于卵泡的复杂功能，卵泡有效地充当离散的微型器官，以促进其所含卵母细胞的生长和成熟。卵母细胞是卵泡的核心，直接被连续的颗粒细胞（GC）层包裹，膜细胞（TC）形成第二层细胞，与卵母细胞和GC结合以构成卵泡单位。虽然分为两组，但GC和TC包含许多亚型。GC根据其在卵泡中的位置进行分类;围绕卵母细胞的GC和邻近基底膜的GC分别被指定为卵泡和壁壁GC，这些亚型显示出独特的转录组特征。TC 具有许多亚型，可提供类固醇生成、代谢和结构支持。内皮细胞、血管周围细胞和免疫细胞在维持正常的卵巢生理学中起着核心作用。卵巢基质不仅可以作为卵泡生长的基质，还可以提供产生TC的祖细胞的来源。卵巢内这种多层细胞亚型复合体使其既是内分泌器官又是生殖器官的功能。
本文提出了一种鉴定和纯化窦卵泡中颗粒细胞、膜细胞、基质细胞、内皮细胞和造血细胞的方案。我们利用该协议分离这些卵巢细胞并使用单细胞测序对其进行分析，然后在不同发育阶段的卵泡中进行特异性染色。该协议提供了一种可复制的简单方法，并且能够对卵巢的生理和病理进行高分辨率分析。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Chemicals, reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic 100x</td>
			<td>Thermo Fisher Scientific</td>
			<td>15240062</td>
			<td>Anti-Anti</td>
		</tr>
		<tr>
			<td>Antifade Mountant solution</td>
			<td>Thermo Fisher Scientific</td>
			<td>&#160;P36930</td>
			<td>ProLong Gold</td>
		</tr>
		<tr>
			<td>Collagenase from&#160;Clostridium histolyticum</td>
			<td>Millipore Sigma</td>
			<td>C 2674</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Thermo Fisher Scientific</td>
			<td>D1306</td>
			<td></td>
		</tr>
		<tr>
			<td>Dispase II, powder</td>
			<td>Thermo Fisher Scientific</td>
			<td>17105041</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Millipore Sigma</td>
			<td>D 2650</td>
			<td>Dimethyl sulfoxide</td>
		</tr>
		<tr>
			<td>DPBS, no calcium, no magnesium</td>
			<td>Thermo Fisher Scientific</td>
			<td>14190144</td>
			<td></td>
		</tr>
		<tr>
			<td>Enzyme Cell Detachment Medium&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>00-4555-56</td>
			<td>Accutase</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum, heat-inactivated</td>
			<td>Thermo Fisher Scientific</td>
			<td>10438026</td>
			<td></td>
		</tr>
		<tr>
			<td>Hanks&#8242; Balanced Salt solution</td>
			<td>Thermo Fisher Scientific</td>
			<td>14175079</td>
			<td>no calcium, no magnesium, no phenol red</td>
		</tr>
		<tr>
			<td>Leibovitz&#8217;s L-15 medium&#160;</td>
			<td>Thermo Fisher Scientific</td>
			<td>11415064</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal Saline</td>
			<td>Quality Biological</td>
			<td>114-055-101</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Millipore Sigma</td>
			<td>S 1888</td>
			<td></td>
		</tr>
		<tr>
			<td>Freezing Medium (100 mL, filtered through a 0.2 micron filter)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>- 69.64 mL of Leibovitz&#39;s L-15</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>- 17.66 mL of fetal bovine serum</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>- 3.42 g of sucrose</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>- 10.65 mL of DMSO</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>- 1 mL of antibiotic-antimycotic</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Lab Plasticware and Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>6-well Clear Flat Bottom Not Treated&#160;</td>
			<td>Corning</td>
			<td>351146</td>
			<td>Falcon</td>
		</tr>
		<tr>
			<td>Cell Strainer 100 &#181;m</td>
			<td>Fisher scientific</td>
			<td>352360</td>
			<td>Corning, Falcon</td>
		</tr>
		<tr>
			<td>Cryovials</td>
			<td>Thermo Fisher Scientific</td>
			<td>377267</td>
			<td>CryoTube 1.8 mL</td>
		</tr>
		<tr>
			<td>Petri dish, D x H 150 mm x 25 mm&#160;</td>
			<td>Millipore Sigma</td>
			<td>CLS430599</td>
			<td>60EA</td>
		</tr>
		<tr>
			<td>Round-Bottom Polystyrene Test Tubes with Cell Strainer Snap Cap, 5 mL</td>
			<td>Fisher scientific</td>
			<td>352235</td>
			<td>Corning, Falcon</td>
		</tr>
		<tr>
			<td>Vacuum Filter/Storage Bottle System, 0.22 &#181;m</td>
			<td>Corning</td>
			<td>431154</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibodies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ANPEP</td>
			<td>BioLegend</td>
			<td>301703</td>
			<td></td>
		</tr>
		<tr>
			<td>CD34</td>
			<td>R&#38;D Systems</td>
			<td>FAB7227A</td>
			<td></td>
		</tr>
		<tr>
			<td>CD45</td>
			<td>BioLegend</td>
			<td>304019</td>
			<td></td>
		</tr>
		<tr>
			<td>CD55</td>
			<td>BioLegend</td>
			<td>311306</td>
			<td></td>
		</tr>
		<tr>
			<td>CD 99</td>
			<td>BioLegend</td>
			<td>371308</td>
			<td></td>
		</tr>
		<tr>
			<td>PVRL</td>
			<td>BioLegend</td>
			<td>340404</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical tools</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>long forceps (~150 mm length)</td>
			<td>Fisherbrand</td>
			<td>12-000-128</td>
			<td>Fisher Scientific</td>
		</tr>
		<tr>
			<td>medium forceps (~110 mm length)</td>
			<td>Fisherbrand</td>
			<td>12-000-157</td>
			<td>Fisher Scientific</td>
		</tr>
		<tr>
			<td>number 21 scalpel</td>
			<td>Andwin Scientific&#160;</td>
			<td>EF7281H</td>
			<td>Fisher Scientific</td>
		</tr>
		<tr>
			<td>number 11 scalpel</td>
			<td>Andwin Scientific</td>
			<td>&#160;FH/CX7281A</td>
			<td>Fisher Scientific</td>
		</tr>
		<tr>
			<td>sharp fine curved scissors</td>
			<td>Roboz Surgical</td>
			<td>RS-5881</td>
			<td></td>
		</tr>
		<tr>
			<td>Instruments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>FACSJazz Flourescence activated cell sorter</td>
			<td>BD</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LSM 710 META Confocal microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

extraction, labeling, and purification of lineage-specific cells from human antral follicles

卵母细胞的活化、生长、发育和成熟是一个复杂的过程，不仅在卵巢的多种细胞类型之间协调，而且在下丘脑/垂体/卵巢回路内的多个控制点之间协调。在卵巢内，多种特化的细胞类型与卵巢卵泡内的卵母细胞密切相关。这些细胞的生物学在后期阶段得到了很好的描述，当它们很容易作为辅助生殖治疗的副产品回收时。然而，由于人类卵巢组织的稀缺和接受辅助生殖治疗的患者对卵巢的访问有限，对直接从卵巢分离的小窦卵泡的深入分析并不常见。 
这些用于处理整个卵巢以冷冻保存皮质条的方法，同时鉴定/分离卵巢驻留细胞，能够对窦卵泡发育的早期阶段进行高分辨率分析。我们展示了通过酶促处理窦卵泡并分离颗粒、膜细胞、内皮细胞、造血细胞和基质细胞来分离离散细胞类型的方案。从各种大小和发育阶段的窦卵泡中分离细胞，可以全面分析驱动卵泡生长和卵巢生理学的细胞和分子机制，并提供可以在 体外 培养的活细胞来源，以概括卵泡微环境。

我们从卵巢表面分离卵泡并对其进行酶处理，以分离GC以及窦腔周围的膜和基质细胞。收集细胞，并通过FACS将细胞组分从窦卵泡（直径范围在0.5mm至4mm之间）分选至>95%纯度（图1）。
为了标记和纯化人窦卵泡内独特的细胞组分，我们将酶消化与流式细胞术分选相结合。我们之前发现了特异性标记卵泡驻留部分的表面蛋白2。我们已经表明...

Watch this Scientific Journal Video about Extraction, Labeling, and Purification of Lineage-Specific Cells from Human Antral Follicles at JoVE.com

Extraction, Labeling, and Purification of Lineage-Specific Cells from Human Antral Follicles

在这里，我们提出了从窦卵泡中鉴定和纯化卵巢细胞的方案。我们详细说明了处理整个卵巢以冷冻保存皮质条的方法，同时还收获完整的窦卵泡，这些卵泡经过酶处理以释放多种卵泡常驻细胞类型，包括颗粒细胞、膜细胞、内皮细胞、造血细胞和基质细胞。

从人窦卵泡中提取、标记和纯化谱系特异性细胞

The activation, growth, development, and maturation of oocytes is a complex process that is coordinated not just between multiple cell types of the ovary ...

这里展示的方案能够标记和分离从早期窦阶段开始存在于卵巢卵泡中的特定细胞谱系，从而实现高分辨率分析和培养。这种高度精确的方法利用荧光团偶联抗体的组合来靶细胞表面标志物，这些标志物在离散的颗粒、基质和膜谱系中稳定且特异性地表达。首先将卵巢放入无菌培养皿中，然后倒入冷却培养基以滋润组织，确保卵巢半浸没在培养基中。移除任何缝合线或其他材料，并将残留的周围组织从卵巢中解剖出来。要分离窦卵泡，请识别单个卵泡并选择健康的卵泡。排除任何充满血液、深色或不对称的卵泡。使用手术刀从卵巢中分离选定的窦卵泡，保持胃窦完整并切除包围卵泡的皮质组织。将每个切除的完整窦卵泡放入六孔板的一个孔中。使用放置在培养皿下方的尺子，确定卵泡直径以进行描述。使用手术刀将完整的窦卵泡一分为二以评估窦腔，并加入6毫升酶细胞分离培养基，然后将板在加湿的培养箱中孵育10分钟。孵育后，加入6毫升含有20%胎牛血清或FBS的可用培养基，并用P-1000微量移液器重复剧烈移液于平分卵泡上，与培养基冲洗。接下来，收集介质并将其通过100毫米过滤器。将过滤后的上清液在300G和4摄氏度下离心三分钟后，吸出上清液。将剩余的组织置于每毫升100单位胶原酶和每毫升1单位分散酶的混合物中，在孵育30分钟前稀释在平衡的盐水溶液中。通过在 10 分钟和 20 分钟后研磨和剧烈移液两次来机械破坏组织。完成后，通过P-1000微量移液器吸头恢复组织并通过移液冲洗，并通过100毫升过滤器过滤。将应变的组织在300G和4摄氏度下离心三分钟，然后将富含膜细胞和基质细胞的细胞沉淀放在冰上进行标记和FACS。为了鉴定颗粒细胞和膜细胞，将细胞沉淀重悬于含有直接偶联抗体的封闭溶液中，并在 4 摄氏度下孵育 10 分钟。洗涤和离心细胞后，将细胞重悬于含有4-6二脒基-2-苯基吲哚或DAPI的FACS缓冲液中，并通过FACS仪器运行细胞悬液以捕获少量细胞，使用荧光团偶联抗体的荧光强度进行门控。将卵巢的其余部分一分为二。使用弯曲的细剪刀和手术刀切除髓质，避免损伤卵巢皮层。通过轻轻刮擦继续处理和变薄卵巢皮层，直到保留最少的髓质。沿卵巢长度将皮质组织分成两到三毫米宽的条带。对于卵巢组织冷冻，将一条皮质条放入每个含有冷冻溶液的冷冻小瓶中。在4摄氏度的旋转板上摇动装有皮质条的低温小瓶20分钟。通过用短暂浸入液氮中的棉头接触每个冷冻小瓶来形成冰晶。收集卵巢细胞，并将细胞组分从窦卵泡FACS分选至纯度超过95%。随着窦卵泡尺寸的增加，PVRL1或凝集素中CD99特异性标记的颗粒细胞越来越多地局限于颗粒卵巢细胞区室。结果发现，针对内分泌蛋白或CD55的抗体特异性地划分了所有基质细胞和膜细胞，丙氨酸氨基肽酶由雄激素膜细胞特异性表达。用针对CD99的抗体标记细胞以鉴定颗粒细胞，并使用抗CD45的抗体来排除造血细胞。针对PVRL1的抗体将颗粒细胞群分离成卵泡和壁室。在用胶原酶分散酶酶消化后，用CD55，CD34和丙氨酸氨基肽酶抗体标记的细胞制剂能够捕获所有基质或膜细胞或雄激素卵泡细胞，而内皮细胞从膜细胞群中排除。在初始卵泡分离期间，通过在卵泡边界周围的安全边缘内切割，特别注意避免破坏卵泡的窦。在该程序的下游，细胞可用于分子或诊断分析，或者用于体外培养以进行毒理学研究，实验分析或卵泡微环境的概括。迄今为止，这些方法被用于模拟涉及卵巢的内分泌疾病，并鉴定卵泡相关细胞的新亚群。

Research

JoVE Journal

Biology

Human ES cells: Starting Culture from Frozen Cells

人类胚胎干细胞：启动文化冷冻细胞

Here we demonstrate how our lab begins a HuES human embryonic stem cell line culture from a frozen stock.  First, a one to two day old ten cm plate of  ...

科研

生物学

In vitro Labeling of Human Embryonic Stem Cells for Magnetic Resonance Imaging

在人类胚胎干细胞体外培养的标签用于磁共振成像

Human embryonic stem cells (hESC) have demonstrated the ability to restore the injured myocardium. Magnetic resonance imaging (MRI) has emerged as one of ...

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

无有机溶剂和醋酸，蛋白质与考马斯亮蓝G - 250凝胶染色

In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents ...

Purification of Mitochondria from Yeast Cells

从酵母细胞线粒体的纯化

Mitochondria are the main site of ATP production during aerobic metabolism in eukaryotic non-photosynthetic cells1. These complex organelles also play ...

Extraction and Analysis of Cortisol from Human and Monkey Hair

提取与分析皮质醇从人与猴的毛

The stress hormone cortisol (CORT) is slowly incorporated into the growing hair shaft of humans, nonhuman primates, and other mammals. We developed and ...

Efficient Production and Purification of Recombinant Murine Kindlin-3 from Insect Cells for Biophysical Studies

高效的生产和纯化重组小鼠Kindlin-3从昆虫细胞生物物理研究

Kindlins are essential coactivators, with talin, of the cell surface receptors&#160;integrins and also participate in integrin outside-in signalling, and ...

Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker

带谱系特异标记的间充质细胞脂肪分化电位的可视化与量化

Several dyes are currently available for use in detecting differentiation of mesenchymal cells into adipocytes. Dyes, such as Oil Red O, are cheap, easy ...

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current

分离高质量鼠心房和心室肌细胞，同时测量 Ca2+ 瞬变和 L 型钙电流

Mouse models play a crucial role in arrhythmia research and allow studying key mechanisms of arrhythmogenesis including altered ion channel function and ...

Tissue Collection and RNA Extraction from the Human Osteoarthritic Knee Joint

从人骨关节炎膝关节收集和RNA提取

Osteoarthritis (OA) is a chronic and degenerative joint disease most often affecting the knee. As there is currently no cure, total knee arthroplasty ...

Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria

Scalable Isolation and Purification of Extracellular Vesicles from Escherichia coli and Other Bacteria

Diverse bacterial species secrete ~20-300 nm extracellular vesicles (EVs), comprised of lipids, proteins, nucleic acids, glycans, and other molecules ...