Name: identification and isolation of burst-forming unit and colony-forming unit erythroid progenitors from mouse tissue by flow cytometry
Uploaded: 2022-11-04T14:00:00
Description: Watch this Scientific Journal Video about Identification and Isolation of Burst-Forming Unit and Colony-Forming Unit Erythroid Progenitors from Mouse Tissue by Flow Cytometry at JoVE.com

这项工作得到了NIH拨款R01DK130498，R01DK120639和R01HL141402的支持。

所有实验均按照动物协议A-1586进行，并202200017马萨诸塞大学陈医学院机构动物护理和使用委员会批准。
注意：此处详细介绍了两种方案：首先，流式细胞术分析（第1节），然后是流式细胞术分选的方案调整（第2节）。以下方案使用具有10个通道的流式细胞仪/分选仪。 表 1 中提供了一个示例设置，如步骤 1.14.5 中所述。也可以仅使用九个通道运行此协议;请参阅 <s...

<ol>
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R., Socolovsky, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stat5+signaling+specifies+basal+versus+stress+erythropoietic+responses+through+distinct+binary+and+graded+dynamic+modalities.">Stat5 signaling specifies basal versus stress erythropoietic responses through distinct binary and graded dynamic modalities.</a> PLoS Biology. 10 (8), 1001383 (2012).</li><li>Koulnis, 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=Contrasting+dynamic+responses+in+vivo+of+the+Bcl-xL+and+Bim+erythropoietic+survival+pathways.">Contrasting dynamic responses in vivo of the Bcl-xL and Bim erythropoietic survival pathways.</a> Blood. 119 (5), 1228-1239 (2012).</li><li>Shearstone, J. 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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=Elucidation+of+the+phenotypic,+functional,+and+molecular+topography+of+a+myeloerythroid+progenitor+cell+hierarchy.">Elucidation of the phenotypic, functional, and molecular topography of a myeloerythroid progenitor cell hierarchy.</a> Cell Stem Cell. 1 (4), 428-442 (2007).</li><li>Dolznig, 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=Expansion+and+differentiation+of+immature+mouse+and+human+hematopoietic+progenitors.">Expansion and differentiation of immature mouse and human hematopoietic progenitors.</a> Methods in Molecular Medicine. 105, 323-344 (2005).</li><li>Li, 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=Isolation+and+transcriptome+analyses+of+human+erythroid+progenitors:+BFU-E+and+CFU-E.">Isolation and transcriptome analyses of human erythroid progenitors: BFU-E and CFU-E.</a> Blood. 124 (24), 3636-3645 (2014).</li><li>Zhang, J., Socolovsky, M., Gross, A. 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直接从高纯度新鲜组织中前瞻性分离BFU-e和CFU-e祖细胞的能力以前是研究人员所没有的。我们的新方法使用scRNAseq和细胞命运测定1，27进行了验证，现在提供了实现这一目标的工具。
成功执行排序和分析方案有许多关键点。首先，细胞需要以900 x g 的速度旋转，以防止低密度细胞的损失。CFU-e阶段的许多细胞是大的低密度细胞?...

红细胞生成可分为两个主要阶段：早期红细胞生成和红系末端分化（图1）1，2，3。在早期红细胞生成中，造血干细胞致力于红系并产生早期红系祖细胞，这些祖细胞在 1970 年代首次根据其在半固体培养基中的集落形成潜力被鉴定4，5，6，7，8，9.从广义上讲，红系祖细胞分为两类：早期祖细胞，每个祖细胞产生“爆发”（较小红细胞簇的大集合），称为“爆发形成单位红细胞”或BFU-e4，5，6;和它们的后代，每个形成一个单一的小红细胞簇或集落，称为“集落形成单位红细胞”或CFU-e7，8，9。BFU-e和CFU-e尚未表达终末红系基因，并且在形态上不可识别。经过多次自我更新或扩增细胞分裂后，CFU-e经历转录开关，其中诱导珠蛋白等红系基因，从而转变为红系末端分化（ETD）1，10。在ETD期间，成红细胞在剜除形成网织红细胞之前经历三到五次成熟细胞分裂，网织红细胞成熟为红细胞。
终末分化期间的成红细胞最初根据其形态分为原成红细胞、嗜碱性、多色和正色。流式细胞术的出现允许基于细胞大小（通过前向散射，FSC测量）和两个细胞表面标记CD71和Ter119 11，12，13进行前瞻性分选和分离（图1）。这种和类似的流式细胞术方法14彻底改变了ETD分子和细胞方面的研究，允许在体内和体外对成红细胞进行发育阶段特异性分析10，15，16，17，18，19，20。CD71/Ter119方法现在常规用于红系前体的分析。
直到最近，研究人员还缺乏一种类似的、可访问的流式细胞术方法，用于从小鼠组织中直接、高纯度地前瞻性分离CFU-e和BFU-e。相反，研究人员使用了流式细胞术策略，仅分离出这些祖细胞的一小部分，通常是在存在在同一流式细胞术亚群中共同纯化的非红系细胞的情况下21。因此，BFU-e和CFU-e的研究仅限于体外分化系统，这些系统从早期骨髓祖细胞衍生和扩增BFU-e和CFU-e。然后可以在这些富含红系祖细胞的培养物中应用流式细胞术策略来区分CFU-e和BFU-e22，23。另一种方法是利用胎儿CFU-e和BFU-e，它们在妊娠中期小鼠胎儿肝脏的Ter119阴性部分中高度富集10，24，25。然而，这两种方法都不允许研究体内生理状态下的成年BFU-e和CFU-e。当回想一下，基于集落形成测定，这些细胞分别以0.025%和0.3%的频率存在于成人骨髓中时，可以理解挑战的严重性6。
这里描述的方案是一种新颖的流式细胞术方法，基于新鲜收获的Kit+小鼠骨髓细胞的单细胞转录组学分析（Kit由骨髓的所有早期祖细胞群表达）1。我们的方法包含一些已被Pronk等人使用的细胞表面标记物21，26。单细胞转录组用于确定识别红系和其他早期造血祖细胞的细胞表面标志物的组合（图2）。具体而言，谱系阴性（Lin-）Kit+细胞的CD55+部分可以细分为五个群体，其中三个产生红系轨迹的连续片段（图2）。通过分选确认这些群体中每个群体的转录组身份，然后进行scRNAseq并将分选的单细胞转录组投影回原始转录组图谱（五个群体中每个群体和整个骨髓数据集的基因表达可以在 https://kleintools.hms.harvard.edu/paper_websites/tusi_et_al/index.html 中探索）1.使用传统的集落形成测定（图2）以及新颖的高通量单细胞命运测定1，27确认了每个群体的细胞命运潜力。这些分析表明，新型流式细胞术方法可高纯度分离新鲜成人骨髓和脾脏的所有BFU-e和CFU-e祖细胞。具体而言，群体 1 （P1） 仅包含 CFU-e 而不包含其他造血祖细胞，群体 2 （P2） 包含骨髓的所有 BFU-e 祖细胞和少量 CFU-e，但没有其他祖细胞1。下面的详细方案通过注射盐水或促红细胞生成激素促红细胞生成素（Epo）的小鼠的示例实验进一步说明。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.5 M EDTA, pH 8.0</td>
			<td>Life Technologies</td>
			<td>15575020</td>
			<td></td>
		</tr>
		<tr>
			<td>1000 &#181;L large orifice tips</td>
			<td>USA sceintific</td>
			<td>1011-9000</td>
			<td></td>
		</tr>
		<tr>
			<td>Alexa Fluor 647 anti-mouse CD55 (DAF) Antibody</td>
			<td>BioLegend</td>
			<td>131806</td>
			<td></td>
		</tr>
		<tr>
			<td>APC/Cyanine7 anti-mouse CD117 (c-kit) Antibody</td>
			<td>BioLegend</td>
			<td>105826</td>
			<td></td>
		</tr>
		<tr>
			<td>Biotin-CD11b</td>
			<td>BD Biosciences</td>
			<td>557395</td>
			<td>M1/70 (clone)</td>
		</tr>
		<tr>
			<td>Biotin-CD19</td>
			<td>BD Biosciences</td>
			<td>553784</td>
			<td>1D3 (clone)</td>
		</tr>
		<tr>
			<td>Biotin-CD4</td>
			<td>BD Biosciences</td>
			<td>BDB553045</td>
			<td>RM4-5 (clone)</td>
		</tr>
		<tr>
			<td>Biotin-CD8a</td>
			<td>BD Biosciences</td>
			<td>BDB553029</td>
			<td>53-6.7 (clone)</td>
		</tr>
		<tr>
			<td>Biotin-F4/80</td>
			<td>Biolegend</td>
			<td>123106</td>
			<td>BM8 (clone)</td>
		</tr>
		<tr>
			<td>Biotin-Ly-6G and Ly-6C</td>
			<td>BD Biosciences</td>
			<td>553125</td>
			<td>RB6-8C5 (clone)</td>
		</tr>
		<tr>
			<td>Biotin-TER-119</td>
			<td>BD Biosciences</td>
			<td>553672</td>
			<td>TER-119 (clone)</td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma aldritch</td>
			<td>A1470</td>
			<td></td>
		</tr>
		<tr>
			<td>Brilliant Violet 421 anti-human/mouse CD49f Antibody</td>
			<td>BioLegend</td>
			<td>313624</td>
			<td></td>
		</tr>
		<tr>
			<td>Brilliant Violet 605 anti-mouse CD41 Antibody</td>
			<td>BioLegend</td>
			<td>133921</td>
			<td></td>
		</tr>
		<tr>
			<td>Brilliant Violet 650 anti-mouse CD150 (SLAM) Antibody</td>
			<td>BioLegend</td>
			<td>115931</td>
			<td></td>
		</tr>
		<tr>
			<td>BUV395 Rat Anti-Mouse TER-119/Erythroid Cells</td>
			<td>BD Biosciences</td>
			<td>563827</td>
			<td></td>
		</tr>
		<tr>
			<td>ChromPure Rabbit IgG, whole molecule</td>
			<td>Jackson ImmunoResearch Laboratories</td>
			<td>011-000-003</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI (4&#39;,6-Diamidino-2-Phenylindole, Dihydrochloride)</td>
			<td>Life Technologies</td>
			<td>D1306</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital DIVA hardware and software for LSR II</td>
			<td>BD Biosciences</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>FITC anti-mouse F4/80 Antibody</td>
			<td>BioLegend</td>
			<td>123108</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Rat Anti-CD11b</td>
			<td>BD Biosciences</td>
			<td>557396</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Rat Anti-Mouse CD19</td>
			<td>BD Biosciences</td>
			<td>553785</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Rat Anti-Mouse CD4</td>
			<td>BD Biosciences</td>
			<td>553047</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Rat Anti-Mouse CD8a</td>
			<td>BD Biosciences</td>
			<td>553031</td>
			<td></td>
		</tr>
		<tr>
			<td>FITC Rat Anti-Mouse Ly-6G and LY-6C</td>
			<td>BD Biosciences</td>
			<td>553127</td>
			<td></td>
		</tr>
		<tr>
			<td>FlowJo software&#160;</td>
			<td>FlowJo</td>
			<td>version 10</td>
			<td>Flow cytometer analysis software</td>
		</tr>
		<tr>
			<td>LSR II digital multiparameter flow cytometer analyzer</td>
			<td>BD Biosciences</td>
			<td></td>
			<td>Flow cytometer&#160;</td>
		</tr>
		<tr>
			<td>NewlineNY Stainless Steel Hand Masher &#38; Bowl, Mortar and Pestle Set</td>
			<td>Amazon</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Normal rat serum</td>
			<td>Stem Cell Technologies</td>
			<td>13551</td>
			<td></td>
		</tr>
		<tr>
			<td>PE anti-mouse CD105 Antibody</td>
			<td>BioLegend</td>
			<td>120408</td>
			<td></td>
		</tr>
		<tr>
			<td>PE/Cyanine7 anti-mouse CD71 Antibody</td>
			<td>BioLegend</td>
			<td>113812</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline, 10x Solution</td>
			<td>Fisher scientific</td>
			<td>BP3994</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptavidin Nanobeads</td>
			<td>BioLegend</td>
			<td>480016</td>
			<td>Magnetic beads</td>
		</tr>
	</tbody>
</table>

identification and isolation of burst-forming unit and colony-forming unit erythroid progenitors from mouse tissue by flow cytometry

早期的红系祖细胞最初由其在体外的集落形成潜力定义，并分为爆发形成和集落形成“单位”，称为BFU-e和CFU-e。直到最近，还没有从新鲜分离的成年小鼠骨髓中直接前瞻性和完全分离纯BFU-e和CFU-e祖细胞的方法。为了解决这一差距，分析了小鼠骨髓的单细胞RNA-seq（scRNAseq）数据集，以表达编码细胞表面标志物的基因。该分析与细胞命运测定相结合，允许开发一种新的流式细胞术方法，该方法可鉴定并允许分离小鼠骨髓或脾脏中BFU-e和CFU-e祖细胞的完整和纯亚群。这种方法还可以识别其他祖细胞亚群，包括富集碱性粒细胞/肥大细胞和巨核细胞潜力的亚群。该方法包括用针对Kit和CD55的抗体标记新鲜骨髓或脾脏细胞。然后，表达这两种标记的祖细胞细分为五个主要群体。群体 1（P1 或 CFU-e，Kit+ CD55+ CD49f 中/低 CD105 中/高 CD71 中/高）包含所有 CFU-e 祖细胞，可进一步细分为 P1-低（CD71 med CD150高）和 P1-hi（CD71高 CD150低），分别对应于早期和晚期 CFU-e; 群体 2（P2 或 BFU-e，Kit+ CD55+ CD49f 中/低 CD105 中/高 CD71低 CD150高）包含所有 BFU-e 祖细胞;群体 P3（P3、试剂盒+ CD55+ CD49f 中/高 CD105中/低 CD150 低 CD41低）富集用于嗜碱性粒细胞/肥大细胞祖细胞;群体 4（P4，Kit+ CD55+ CD49f 中/高 CD105中/低 CD150高 CD41+）富集巨核细胞祖细胞;和群体 5（P5、Kit+ CD55+ CD49f 中/高 CD105 中/低 CD150高 CD41-）包含具有红细胞、嗜碱性粒细胞/肥大细胞和巨核细胞潜力 （EBMP） 和红细胞/巨核细胞/嗜碱性粒细胞偏向的多电位祖细胞 （MPP） 的祖细胞。这种新方法在分析红系和其他造血祖细胞时可以提高精度，并且还可以参考每个流式细胞术定义的群体的转录组信息。

该协议描述了一种流式细胞术方法，用于鉴定新鲜收获的骨髓和脾脏细胞中的BFU-es和CFU-es。它首先从小鼠身上收获新鲜的BM和脾脏，并立即将组织放在冰上。所有程序均在低温下进行，以保持细胞活力。用“谱系”抗体混合物标记细胞，该混合物允许排除所有表达分化血系标志物的细胞（FITC-Lin鸡尾酒，表 3，在流式细胞术分析的情况下;或磁珠富集， 表4，用于谱系阴性细...

Watch this Scientific Journal Video about Identification and Isolation of Burst-Forming Unit and Colony-Forming Unit Erythroid Progenitors from Mouse Tissue by Flow Cytometry at JoVE.com

Identification and Isolation of Burst-Forming Unit and Colony-Forming Unit Erythroid Progenitors from Mouse Tissue by Flow Cytometry

在这里，我们描述了一种新的流式细胞术方法，用于直接从新鲜小鼠骨髓和脾脏中前瞻性分离早期爆发形成单位红系（BFU-e）和集落形成单位红系（CFU-e）祖细胞。该协议基于单细胞转录组学数据开发，是第一个以高纯度分离所有组织红系祖细胞的方案。

流式细胞术从小鼠组织中鉴定和分离爆破形成单元和集落形成单元红系祖细胞

Early erythroid progenitors were originally defined by their colony-forming potential in vitro and classified into burst-forming and colony-forming ...

该协议允许直接从新鲜收获的小鼠组织（如骨髓和脾脏）中鉴定和分离BFU-E和CFU-E红系祖细胞。使用这种技术，我们可以分离出纯粹的红系祖细胞群，这在以前的流动方案中是不可能的。这种方法通过允许在许多下游实验中分离祖细胞，极大地增强了我们研究红系疾病小鼠模型的能力。首先，将新鲜解剖的股骨和胫骨直接放入冷染色缓冲液或SB-5中，并将管子放在冰上，直到准备好提取骨髓。将干净的消毒砂浆放在桶中的冰上，然后将骨头转移到研钵中。使用解剖剪刀，剪掉每根骨头的大约 1 毫米末端。使用配备含有SB-5的26号针头的3毫升注射器将骨髓从骨头中冲洗出来并将其收集到研钵中，每次使用新鲜缓冲液重复该过程3至5次，直到骨骼呈现无色。通过100微米网格过滤冲洗的骨髓，并将细胞收集在放置在冰上的50毫升离心管中。接下来，用研钵和研杵将冲洗过的骨头压碎在 5 到 10 毫升的 SB-5 中。过滤碎骨的混合物，并通过100微米细胞过滤器缓冲液，与先前收集的细胞汇集。在放置在冰上的空50毫升离心管上设置100微米网状过滤器。将单个脾脏放在网状过滤器上，并在脾脏中加入 0.5 至 1 毫升 SB-5，以确保其不会干燥。使用3或5毫升注射器柱塞的橡胶侧，将脾脏捣碎。添加更多的SB-5，一次5毫升，并继续捣碎，直到所有细胞都收集在管中。通过使用大孔尖端用 2 毫升 SB-5 上下移液收集的细胞来制备单细胞悬液。通过将除 FMO 同名抗体以外的所有抗体添加到 FACS 管中的细胞悬液中，对荧光减去 1 或 FMO 对照进行染色。要对单色对照进行染色，请将单一抗体添加到FACS管中的细胞悬液中。以 3000 RPM 涡旋每个控制管 2 至 3 秒，然后在 4 摄氏度下在黑暗中摇摆 2.5 小时。孵育后，用SB-5洗涤细胞，并用SB-5将细胞悬液的体积补足至4毫升。在4摄氏度下以900g旋转细胞10分钟并吸出上清液。将未染色的和所有单色对照重新悬浮在 300 微升 SB-5 中，并通过 40 微米过滤网过滤到新的 FACS 管中。通过在SB-5中以1比10， 000的比例稀释DAPI SB-5来制备DAPI SB-5溶液。将FMOS重新悬浮在1.8毫升DAPI SB-5中，将DAPI单色对照重新悬浮在300微升DAPI SB-5中，然后通过40微米过滤网将它们过滤到新的FACS管中。将抗体预混液添加到每个样品中后，以 3000 rpm 的速度涡旋试管 2 至 3 秒，然后在黑暗中在 4 摄氏度下在摇摆条件下孵育 2.5 小时。如前所述洗涤细胞后，将样品重新悬浮在 1.8 毫升 DAPI SB-5 中，并通过 40 微米过滤网过滤到新的 FACS 管中，并使用膀胱计分析样品。使用前向散射根据大小去除碎片，然后使用前向散射高度与前向散射区域直方图排除细胞聚集并选择单个细胞。使用侧向散射高度与侧向散射区域直方图重复排除。通过门控DAPI的阳性细胞来排除死细胞。选择谱系阴性TUR一个19阴性试剂盒阳性细胞。并从中选择一个表达CD 55的亚群。根据CD 49 F和CD 1 0 5的表达，将谱系阴性TUR 19，阴性试剂盒，阳性，CD 55阳性细胞细分为2个主要群体，P六和P七。现在基于CD 150和CD 41的表达，将P 6进一步细分为P 3，P 4和P 5。同样，基于CD 150和CD 71的表达，将P 7进一步细分为BFUE，早期CFUE和晚期CFUE。在本研究中，从新鲜收获的骨髓和脾脏细胞中鉴定出爆裂和集落形成单位。在小鼠中注射促红细胞生成素或载体72小时后，促红细胞生成素刺激显示收获的骨髓和脾脏细胞中早期和晚期集落形成单位群体P1低和P 1高扩张。与载体对照相比，骨髓和脾脏祖细胞对体内促红细胞生成素的反应也显示出早期和晚期集落形成单位群体P 1低和P 1高细胞数量的细胞。最重要的是在用抗体标记细胞之前产生单细胞悬液。这可以通过反复上下移液以及在缓冲液中加入EDTA来实现。纯化的祖细胞可用于任何下游细胞和分子分析。例如，单细胞转录组学、atesque、集落测定和生物化学。这项技术帮助我们测试了单细胞RNA-seq实验的预测。

Research

JoVE Journal

Developmental Biology

Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays

Characterization of Thymic Settling Progenitors in the Mouse Embryo Using In Vivo and In Vitro Assays

胸腺表征祖定居在小鼠胚胎使用<em&gt;在体内</em&gt;和<em&gt;在体外</em&gt;测定

Characterizing thymic settling progenitors is important to understand the pre-thymic stages of T cell development, essential to devise strategies for T ...

科研

发育生物学

Step-specific Sorting of Mouse Spermatids by Flow Cytometry

步特定老鼠的精子细胞分选流式细胞仪

The differentiation of mouse spermatids is one critical process for the production of a functional male gamete with an intact genome to be transmitted to ...

Identification Of Erythromyeloid Progenitors And Their Progeny In The Mouse Embryo By Flow Cytometry

通过流式细胞术鉴定小鼠胚胎中的红细胞生成素及其后代

Macrophages are professional phagocytes from the innate arm of the immune system. In steady-state, sessile macrophages are found in adult tissues where ...

Isolating and Analyzing Cells of the Pancreas Mesenchyme by Flow Cytometry

隔离并通过流式细胞仪分析胰腺间充质细胞

The pancreas is comprised of epithelial cells that are required for food digestion and blood glucose regulation. Cells of the pancreas microenvironment, ...

Expansion and Adipogenesis Induction of Adipocyte Progenitors from Perivascular Adipose Tissue Isolated by Magnetic Activated Cell Sorting

通过磁性活化细胞分选分离的血管周围脂肪组织的脂肪细胞祖细胞的扩增和脂肪生成诱导

Expansion of Perivascular Adipose Tissue (PVAT), a major regulator of vascular function through paracrine signaling, is directly related to the ...

A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry

通过机械组织分离和流式细胞术对哺乳动物雄性生殖细胞多特异性纯化的标准化方法

Fluorescence-activated cell sorting (FACS) has been one of the methods of choice to isolate enriched populations of mammalian testicular germ cells. ...

Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice

小鼠骨髓基质细胞与骨破骨祖的分离、培养及分化

Bone marrow stromal cells (BMSCs) constitute a cell population routinely used as a representation of mesenchymal stem cells in vitro. They reside within ...

Isolation and Characterization of Cardiac Mesenchymal Stromal Cells from Endomyocardial Bioptic Samples of Arrhythmogenic Cardiomyopathy Patients

致心律失常心肌病患者心内膜心肌活检样间质基质细胞的分离与鉴定

A normal adult heart is composed of several different cell types, among which cardiac mesenchymal stromal cells represent an abundant population. The ...

Direct Lineage Reprogramming of Adult Mouse Fibroblast to Erythroid Progenitors

成人小鼠成纤维细胞对红细胞生成子细胞的直接线重编程

Erythroid cell commitment and differentiation proceed through activation of a lineage-restricted transcriptional network orchestrated by a group of cell ...

Real-time Live-cell Flow Cytometry to Investigate Calcium Influx, Pore Formation, and Phagocytosis by P2X7 Receptors in Adult Neural Progenitor Cells

P2X7 受体实时活细胞流式细胞仪检测成人神经祖细胞中的钙影响、毛孔形成和吞噬作用

Live-cell flow cytometry is increasingly used among cell biologists to quantify biological processes in a living cell culture. This protocol describes a ...