Name: an efficient method for directed hepatocyte-like cell induction from human embryonic stem cells
Uploaded: 2021-05-06T15:00:00
Description: Watch this Scientific Journal Video about An Efficient Method for Directed Hepatocyte-Like Cell Induction from Human Embryonic Stem Cells at JoVE.com

这项工作得到了中国国家自然科学基金委员会（第81870432号，81570567 X.L.Z.）（第81571994号）的支持：中国广东省自然科学基金（第2020A1515010054号，中华人民共和国），李嘉诚汕头大学基金会（第2020A15015010054号）。L1111 2008 至 P.N.S.）。我们要感谢汕头大学医学院的林士丹教授的有益建议。

1. 干细胞维护
注：下文所述的细胞维护协议适用于在附着单层中维护的 hES03 细胞系。对于本手稿中的所有以下协议，细胞应在生物安全柜下处理。
<ol><li>通过稀释5倍补充介质到mTesR基础介质，准备1xmTesR干细胞培养介质。</li>
	<li>通过在冰面上稀释 5 毫升符合 hESC 资格的矩阵凝胶，用 5 毫升的 DMEM/F12 来制备 30 倍 hESC 合格的矩阵介质。存储在-20°C。</li>
	<li>通过稀释 33.3 μL 的...

<ol>
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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=PI3K/mTORC2+regulates+TGF-beta/Activin+signalling+by+modulating+Smad2/3+activity+via+linker+phosphorylation.">PI3K/mTORC2 regulates TGF-beta/Activin signalling by modulating Smad2/3 activity via linker phosphorylation.</a> Nature Communications. 6, 7212 (2015).</li><li>Borowiak, 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=Small+molecules+efficiently+direct+endodermal+differentiation+of+mouse+and+human+embryonic+stem+cells.">Small molecules efficiently direct endodermal differentiation of mouse and human embryonic stem cells.</a> Cell Stem Cell. 4, 348-358 (2009).</li><li>Tahamtani, 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=Treatment+of+human+embryonic+stem+cells+with+different+combinations+of+priming+and+inducing+factors+toward+definitive+endoderm.">Treatment of human embryonic stem cells with different combinations of priming and inducing factors toward definitive endoderm.</a> Stem Cells and Development. 22, 1419-1432 (2013).</li><li>Song, 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=Efficient+generation+of+hepatocyte-like+cells+from+human+induced+pluripotent+stem+cells.">Efficient generation of hepatocyte-like cells from human induced pluripotent stem cells.</a> Cell Research. 19, 1233-1242 (2009).</li><li>Sullivan, G. 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=Generation+of+functional+human+hepatic+endoderm+from+human+induced+pluripotent+stem+cells.">Generation of functional human hepatic endoderm from human induced pluripotent stem cells.</a> Hepatology. 51, 329-335 (2010).</li><li>Xia, 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=Human+stem+cell-derived+hepatocytes+as+a+model+for+hepatitis+B+virus+infection,+spreading+and+virus-host+interactions.">Human stem cell-derived hepatocytes as a model for hepatitis B virus infection, spreading and virus-host interactions.</a> Journal of Hepatology. 66, 494-503 (2017).</li><li>Li, 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=Derivation+and+applications+of+human+hepatocyte-like+cells.">Derivation and applications of human hepatocyte-like cells.</a> World Journal of Stem Cells. 11, 535-547 (2019).</li><li>Ogawa, 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=Three-dimensional+culture+and+cAMP+signaling+promote+the+maturation+of+human+pluripotent+stem+cell-derived+hepatocytes.">Three-dimensional culture and cAMP signaling promote the maturation of human pluripotent stem cell-derived hepatocytes.</a> Development. 140, 3285-3296 (2013).</li><li>Kim, D. 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在这里，我们提出了一个分步方法，将 HSC 分为三个阶段。在第一阶段，阿克蒂文A和CHIR99021用于区分氢氟碳化合物到DE。在第二阶段，KO-DMEM和DMSO用于区分DE到肝祖细胞。在第三阶段，HZM加HGF、OSM和氢皮质松21-血氧钠盐用于继续将肝祖细胞分化为HLC。
在使用协议时，需要考虑以下关键步骤。细胞的初始分化密度和中等变化的准确时间是成功分化的重要因素。当我们开始分化时?...

肝脏是一种高度代谢的器官，起着多种作用，包括脱氧、储存糖原、蛋白质的分泌和合成等。各种病原体、药物和羊群可引起肝脏病理变化，影响其功能。肝细胞作为肝脏的主要功能单位，在人工肝脏支持系统和药物毒性消除方面发挥着重要作用。然而，在细胞治疗和肝病研究中，原发性人类肝细胞的资源有限。因此，开发新的功能性人类肝细胞来源是再生医学领域的一个重要研究方向。自1998年建立HESC以来，由于具有优越的分化潜力（可以在合适的环境中分化成各种组织）和高度的自我更新性，为生物关节肝、肝细胞移植甚至肝组织工程提供了理想的来源细胞，因此，hESC得到了广泛的应用。
目前，通过丰富内分泌物6，可以大大提高肝分化效率。在干细胞分泌成内分泌物的系数中，转化生长因子β（TGF-β）信号和WNT信号通路的水平是内分泌形成阶段节点的关键因素。激活高水平的TGF-β和WNT信号可以促进内分泌体7、8的发展。阿克蒂文A是属于TGF-β超级家庭的细胞因子。因此，Activin A广泛应用于人类诱导多能干细胞（hiPSC）和hESC9、10的内皮体诱导。葛兰素史克3是一种血清素-三氨酸蛋白激酶。研究人员发现，CHIR99021是葛兰素史克3®的特异性抑制剂，可以刺激典型的WNT信号，并在一定条件下促进干细胞分化，表明CHIR99021有可能诱导干细胞分化成内分泌物11、12、13。 
在这里，我们报告一种高效和可重复的方法，用于有效诱导 hESC 分化为功能 HHLC。Activin A 和 CHIR99021 的连续添加产生了约 89.7 ±0.8% SOX17 （DE 标记） 阳性细胞。 在体外进一步成熟后，这些细胞表示肝特异性标记，并施加肝细胞状形态（基于血氧林-肌辛染色（H &E））和功能，如吸收花青绿色（ICG）、糖原储存和CYP3活性。结果表明，HESC可以通过这种方法成功分化为成熟的功能HLC，为肝病相关研究和 体外 药物筛选提供依据。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2-Mercaptoethanol</td>
			<td>Sigma</td>
			<td>M7522</td>
			<td>For hepatic progenitor differentiation</td>
		</tr>
		<tr>
			<td>488 labeled goat against mouse IgG</td>
			<td>ZSGB-BIO</td>
			<td>ZF-0512</td>
			<td>For IF,second antibody</td>
		</tr>
		<tr>
			<td>488 labeled goat against Rabbit IgG</td>
			<td>ZSGB-BIO</td>
			<td>ZF-0516</td>
			<td>For IF,second antibody</td>
		</tr>
		<tr>
			<td>Accutase</td>
			<td>Stem Cell Technologies</td>
			<td>7920</td>
			<td>For cell passage</td>
		</tr>
		<tr>
			<td>Activin A</td>
			<td>peproTech</td>
			<td>120-14E</td>
			<td>For definitive endoderm formation</td>
		</tr>
		<tr>
			<td>Anti - Albumin (ALB)</td>
			<td>Sigma-Aldrich</td>
			<td>A6684</td>
			<td>For IF and WB, primary antibody</td>
		</tr>
		<tr>
			<td>Anti - Human Oct4</td>
			<td>Abcam</td>
			<td>Ab19587</td>
			<td>For IF, primary antibody</td>
		</tr>
		<tr>
			<td>Anti - &#945;-Fetoprotein (AFP)</td>
			<td>Sigma-Aldrich</td>
			<td>A8452</td>
			<td>For IF and WB, primary antibody</td>
		</tr>
		<tr>
			<td>Anti -SOX17</td>
			<td>Abcam</td>
			<td>ab224637</td>
			<td>For IF, primary antibody</td>
		</tr>
		<tr>
			<td>Anti-Hepatocyte Nuclear Factor 4 alpha (HNF4&#945;)</td>
			<td>Sigma-Aldrich</td>
			<td>SAB1412164</td>
			<td>For IF, primary antibody</td>
		</tr>
		<tr>
			<td>B-27 Supplement</td>
			<td>Gibco</td>
			<td>17504-044</td>
			<td>For definitive endoderm formation</td>
		</tr>
		<tr>
			<td>BSA</td>
			<td>Beyotime</td>
			<td>ST023-200g</td>
			<td>For cell blocking</td>
		</tr>
		<tr>
			<td>CHIR99021</td>
			<td>Sigma-Aldrich</td>
			<td>SML1046</td>
			<td>For definitive endoderm formation</td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Beyotime</td>
			<td>C1006</td>
			<td>For nuclear staining</td>
		</tr>
		<tr>
			<td>DEPC-water</td>
			<td>Beyotime</td>
			<td>R0021</td>
			<td>For RNA dissolution</td>
		</tr>
		<tr>
			<td>DM3189</td>
			<td>MCE</td>
			<td>HY-12071</td>
			<td>For definitive endoderm formation</td>
		</tr>
		<tr>
			<td>DMEM/F12</td>
			<td>Gibco</td>
			<td>11320-033</td>
			<td>For cell culture</td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma-Aldrich</td>
			<td>D5879</td>
			<td>For hepatic progenitor differentiation</td>
		</tr>
		<tr>
			<td>DPBS</td>
			<td>Gibco</td>
			<td>14190-144</td>
			<td>For cell culture</td>
		</tr>
		<tr>
			<td>GlutaMAX</td>
			<td>Gibco</td>
			<td>35050-061</td>
			<td>For hepatic progenitor differentiation</td>
		</tr>
		<tr>
			<td>H&#38;E staining kit</td>
			<td>Beyotime</td>
			<td>C0105S</td>
			<td>For H&#38;E staining</td>
		</tr>
		<tr>
			<td>Hepatocyte growth factor (HGF)</td>
			<td>peproTech</td>
			<td>100-39</td>
			<td>For hepatocyte differentiation</td>
		</tr>
		<tr>
			<td>HepatoZYME-SFM (HZM)</td>
			<td>Gibco</td>
			<td>17705-021</td>
			<td>For hepatocyte differentiation</td>
		</tr>
		<tr>
			<td>Hydrocortisone-21-hemisuccinate</td>
			<td>Sigma-Aldrich</td>
			<td>H4881</td>
			<td>For hepatocyte differentiation</td>
		</tr>
		<tr>
			<td>Indocyanine</td>
			<td>Sangon Biotech</td>
			<td>A606326</td>
			<td>For Indocyanine staining</td>
		</tr>
		<tr>
			<td>Knock Out DMEM</td>
			<td>Gibco</td>
			<td>10829-018</td>
			<td>For hepatic progenitor differentiation</td>
		</tr>
		<tr>
			<td>Knock Out SR Multi-Species</td>
			<td>Gibco</td>
			<td>A31815-02</td>
			<td>For hepatic progenitor differentiation</td>
		</tr>
		<tr>
			<td>Matrigel hESC-qualified</td>
			<td>Corning</td>
			<td>354277</td>
			<td>For cell culture</td>
		</tr>
		<tr>
			<td>MEM NeAA</td>
			<td>Gibco</td>
			<td>11140-050</td>
			<td>For hepatic progenitor differentiation</td>
		</tr>
		<tr>
			<td>mTesR 5X Supplement</td>
			<td>Stem Cell Technologies</td>
			<td>85852</td>
			<td>For cell culture</td>
		</tr>
		<tr>
			<td>mTesR Basal Medium</td>
			<td>Stem Cell Technologies</td>
			<td>85851</td>
			<td>For cell culture</td>
		</tr>
		<tr>
			<td>Oncostatin (OSM)</td>
			<td>peproTech</td>
			<td>300-10</td>
			<td>For hepatocyte differentiation</td>
		</tr>
		<tr>
			<td>P450 - CYP3A4 (Luciferin - PFBE)</td>
			<td>Promega</td>
			<td>V8901</td>
			<td>For CYP450 activity</td>
		</tr>
		<tr>
			<td>PAS staining kit</td>
			<td>Solarbio</td>
			<td>G1281</td>
			<td>For PAS staining</td>
		</tr>
		<tr>
			<td>Pen/Strep</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td>For cell differentiation</td>
		</tr>
		<tr>
			<td>Peroxidase-Conjugated Goat anti-Mouse IgG</td>
			<td>ZSGB-BIO</td>
			<td>ZB-2305</td>
			<td>For WB,second antibody</td>
		</tr>
		<tr>
			<td>Primary Antibody Dilution Buffer for&#160;Western Blot</td>
			<td>Beyotime</td>
			<td>P0256</td>
			<td>For primary antibody dilution</td>
		</tr>
		<tr>
			<td>ReverTraAce qPCR RT Kit</td>
			<td>TOYOBO</td>
			<td>FSQ-101</td>
			<td>For cDNA Synthesis</td>
		</tr>
		<tr>
			<td>RNAiso Plus</td>
			<td>TaKaRa</td>
			<td>9109</td>
			<td>For RNA Isolation</td>
		</tr>
		<tr>
			<td>RPMI 1640</td>
			<td>Gibco</td>
			<td>11875093</td>
			<td>For definitive endoderm formation</td>
		</tr>
		<tr>
			<td>Skim milk</td>
			<td>Sangon Biotech</td>
			<td>A600669</td>
			<td>For second antibody preparation</td>
		</tr>
		<tr>
			<td>SYBR Green Master Mix</td>
			<td>Thermo Fisher Scientific</td>
			<td>A25742</td>
			<td>For RT-PCR Analysis</td>
		</tr>
		<tr>
			<td>Torin2</td>
			<td>MCE</td>
			<td>HY-13002</td>
			<td>For definitive endoderm formation</td>
		</tr>
		<tr>
			<td>Tween</td>
			<td>Sigma-Aldrich</td>
			<td>WXBB7485V</td>
			<td>For washing buffer preparation</td>
		</tr>
	</tbody>
</table>

an efficient method for directed hepatocyte-like cell induction from human embryonic stem cells

从人类胚胎干细胞中提取的肝细胞样细胞（HMC）的潜在功能对疾病建模和药物筛选应用有很大的希望。如果这里是将 hESC 区分为功能性 HMC 的有效且可重复的方法。建立内皮体血统是区分HLC的关键步骤。通过我们的方法，我们通过在 hESC 分化过程中持续补充 Activin A 和 CHIR99021 来调节关键信号通路，然后生成肝祖细胞，最后在具有完全定义试剂的阶段性方法中使用典型的肝细胞形态学的 HLC。这种方法产生的 hESC 衍生 HLC 表示特定阶段标记（包括白蛋白， HNF4®核受体和陶罗乔拉酸钠共运多肽（NTCP），并表现出与成熟和功能性肝细胞（包括花青绿色染色、糖原储存、血氧林-异丙辛染色和CYP3活性）相关的特殊特性，为肝病研究中基于HLC的应用的发展提供了平台。

HLC 感应的示意图图和每个分化阶段的代表性亮场图像显示在图 1 中。在第一阶段，Activin A和CHIR99021被添加3天，以诱导干细胞形成内皮细胞。在第二阶段，内皮细胞在用分化介质治疗5天后分化成肝祖细胞。在第三阶段，早期肝细胞在HGF和OSM（图1A）中10天后成熟并分化为HLC。在分化的最后阶段，细胞表现出典型的肝细胞表型（细胞是多边形的，分布均匀...

Watch this Scientific Journal Video about An Efficient Method for Directed Hepatocyte-Like Cell Induction from Human Embryonic Stem Cells at JoVE.com

An Efficient Method for Directed Hepatocyte-Like Cell Induction from Human Embryonic Stem Cells

本手稿描述了人类胚胎干细胞（hESC）分化成功能性肝细胞样细胞（HLCs）的详细协议，在hESC分化过程中连续补充Activin A和CHIR99021，以最终内分泌体（DE）。

从人类胚胎干细胞中引导肝细胞样细胞诱导的有效方法

The potential functions of hepatocyte-like cells (HLCs) derived from human embryonic stem cells (hESCs) hold great promise for disease modeling and drug ...

人类胚胎干细胞衍生的肝细胞样细胞在疾病建模和药物筛选中很有用。这是有效诱导人类胚胎干细胞分化成功能性肝细胞样细胞的有效可重复方法。人类胚胎干细胞的无差别状态和适当汇合对于成功分化肝细胞细胞至关重要。为了维持干细胞，涂层无菌的六井组织培养处理板与1毫升1倍的人类胚胎干细胞合格Matrigel每口井，并储存在4摄氏度过夜。在使用前将盘子放在室温下30分钟。在37摄氏度的水浴中解冻冷冻保存的 HESEs 三分钟而不摇晃，然后立即将干细胞输送到含有 4 毫升预热、37 摄氏度 MTESR 介质的 15 毫升离心管中。离心液和吸气超自然，然后轻轻地重新悬浮细胞在一毫升的MTESR介质。从盘子中吸气 DMEM F-12 介质。将细胞播种到六井板中，密度为1×10至5个细胞，在两毫升的MTESR中孵化，并在5%的二氧化碳孵化器中孵化。通过每天用预热的 mTESR 介质替换介质来维护细胞。在大约70%到80%的汇合中，或者当细胞群落开始接触时，通过细胞。对于通过的细胞，吸气介质，并在37摄氏度下用每口酶溶液一毫升孵育细胞5分钟。将细胞通过管道输送到15毫升离心管中，管内含有4毫升DMEM F-12预热至37摄氏度。准备24井板涂上250微升的1X hESC合格的矩阵介质。种子 hESCs 的密度为 mTESR 介质的 500 微升，每口井的密度为 1 到 1.5 倍 10 至第 5 个细胞。将 Activin A 添加到每毫升 100 纳米的最终浓度和 CHIR99021 的最终浓度中，在适当量的预热阶段一分化基本介质中加入三微摩尔。经过三天的分化，分化细胞应表示最终内皮细胞的标记，如FOXA2、SOX17、GATA4、CXCR4和FOXA1。在分化的第三天，吸气介质，替换为第二阶段介质，孵育24小时。在第四天到第八天每隔一天更换一次介质。经过八天的分化，分化细胞应表示肝祖细胞的适当标记，如HNF4α、AFP、TBX3、TTR、ALB、NTCP、CEBPA。在第八天，从细胞中吸气第二阶段介质，代之以第三阶段介质。允许细胞在37摄氏度的二氧化碳孵化器中孵化10天，每隔一天用新添加的分化因子改变介质。经过18天的分化，分化细胞应表示肝细胞的特征标记，如AAT、ALB、TTR、HNF4阿尔法、NTCP、ASGR1、CYP3A4。显示了 hESC 中肝细胞状细胞的示意图图和每个分化阶段的明亮场图像。在第一阶段，ACTIVIN A和CHIR99021被添加三天，以诱导干细胞形成内皮细胞。在第二阶段，内皮细胞在用分化介质治疗五天后分化成肝祖细胞。在第三阶段，10天后，早期肝细胞已经成熟并分化成肝细胞状细胞中的肝细胞生长因子和雄激素。在分化的最后阶段，细胞表现出典型的肝细胞表型。RT-PCR、免疫荧光染色和西性染色用于检测内皮细胞、肝前体细胞和成熟肝细胞的标记物。分化细胞在每个阶段都显示出高表达水平的分化相关基因和蛋白质，如第三天SOS17的内皮标记、第8天的肝前体标记HNF4阿尔法、第14天的法新社和第18天的成熟肝细胞标记ALB。HLC 表现出绿色染色和广泛的细胞质酸转移染色。肝细胞表现出典型的双核形态。CYP3A4的酶活性，即HLC肝脏中必需的代谢CYP，通过酶检测得到证实。将人类胚胎干细胞定位在适当的密度下，并在下一天开始分化至关重要。在第一阶段，轻轻触摸介质，因为细胞容易浮起来。Activin A和其他小分子的结合可以进一步提高分化效率，并在此过程中产生更成熟的肝细胞状细胞。该技术可为探索肝细胞移植、肝组织工程、生物人工肝和疾病建模提供平台。

Research

JoVE Journal

Immunology and Infection

A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells

一种简单而有效的隔离混合原代培养成人肝细胞巨噬细胞的方法

Kupffer cells are liver-specific resident macrophages and play an important role in the physiological and pathological functions of the liver1-3. Although ...

科研

免疫与感染

Optimized Protocol for Efficient Transfection of Dendritic Cells without Cell Maturation

高效转染树突状细胞的无细胞的成熟优化的协议

Dendritic cells (DCs) can be considered sentinels of the immune system which play a critical role in its initiation and response to infection1. Detection ...

HLA-Ig Based Artificial Antigen Presenting Cells for Efficient ex vivo Expansion of Human CTL

HLA-Ig Based Artificial Antigen Presenting Cells for Efficient ex vivo Expansion of Human CTL

HLA - IG基于人工抗原提呈细胞可高效体外扩展人类的CTL

CTL with optimal effector function play critical roles in mediating protection against various intracellular infections and cancer. However, individuals ...

Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes

定向分化诱导性多潜能干细胞向T淋巴细胞

Adoptive cell transfer (ACT) of antigen-specific CD8+  cytotoxic T lymphocytes (CTLs) is a promising treatment for a variety of  malignancies 1. CTLs can  ...

Killer Artificial Antigen Presenting Cells (KaAPC) for Efficient In Vitro Depletion of Human Antigen-specific T Cells

Killer Artificial Antigen Presenting Cells KaAPC for Efficient In Vitro Depletion of Human Antigen-specific T Cells

杀手人工抗原提呈细胞（KaAPC）为有效<em&gt;在体外</em&gt;人抗原特异性T细胞耗竭

Current treatment of T cell mediated autoimmune diseases relies mostly on strategies of global immunosuppression, which, in the long term, is accompanied ...

Derivation of T Cells In Vitro from Mouse Embryonic Stem Cells

Derivation of T Cells In Vitro from Mouse Embryonic Stem Cells

对T细胞推导<em&gt;在体外</em&gt;从小鼠胚胎干细胞

The OP9/OP9-DL1 co-culture system has become a well-established method for deriving differentiated blood cell types from embryonic and hematopoietic ...

An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices

An Ex vivo Model of an Oligodendrocyte-directed T-Cell Attack in Acute Brain Slices

一个<em&gt;体外</em&gt;急性脑片的少突胶质细胞定向T细胞攻击模式

Death of oligodendrocytes accompanied by destruction of neurons and axons are typical histopathological findings in cortical and subcortical grey matter ...

An Optimized Method for Isolating and Expanding Invariant Natural Killer T Cells from Mouse Spleen

从小鼠脾隔离和扩大不变的自然杀伤T细胞的优化方法

The ability to rapidly secrete cytokines upon stimulation is a functional characteristic of the invariant natural killer T (iNKT) cell lineage. iNKT cells ...

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. ...

Efficient Isolation Protocol for B and T Lymphocytes from Human Palatine Tonsils

从人类腭扁桃体B和T淋巴细胞有效隔离协议

Tonsils form a part of the immune system providing the first line of defense against inhaled pathogens. Usually the term &#8220;tonsils&#8221; refers to ...