Name: human pluripotent stem cell based developmental toxicity assays for chemical safety screening and systems biology data generation
Uploaded: 2015-06-17T13:00:00.000+00:00
Duration: 17 min 28 s
Description: Watch this Scientific Journal Video about Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation at JoVE.com

We thank M. Kapitza, Margit Henry, Tamara Rotshteyn, Susan Rohani and Cornelia B&#246;ttinger for excellent technical support. This work was supported by grants from the German Research Foundation (RTG 1331) and the German Ministry for Research (BMBF).

<ol>
	<li>Liu, W. W., Deng, Y. G., Liu, Y., Gong, W. R., Deng, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+Cell+Models+for+Drug+Discovery+and+Toxicology+Studies.">Stem Cell Models for Drug Discovery and Toxicology Studies.</a> Journal of Biochemical and Molecular Toxicology. 27 (1), 17-27 (2013).</li><li>Zuba-Surma, E. K., Jozkowicz, A., Dulak, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+Cells+in+Pharmaceutical+Biotechnology.">Stem Cells in Pharmaceutical Biotechnology.</a> Current Pharmaceutical Biotechnology. 12 (11), 1760-1773 (2011).</li><li>Leist, M., Hartung, T., Nicotera, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dawning+of+a+new+age+of+toxicology.">The dawning of a new age of toxicology.</a> ALTEX. 25 (2), 103-114 (2008).</li><li>Kuegler, P. B., 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=Markers+of+murine+embryonic+and+neural+stem+cells,+neurons+and+astrocytes:+reference+points+for+developmental+neurotoxicity+testing.">Markers of murine embryonic and neural stem cells, neurons and astrocytes: reference points for developmental neurotoxicity testing.</a> ALTEX. 27 (1), 17-42 (2010).</li><li>Yamada, 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=Human+oocytes+reprogram+adult+somatic+nuclei+of+a+type+1+diabetic+to+diploid+pluripotent+stem+cells.">Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells.</a> Nature. , (2014).</li><li>Chambers, S. 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=Highly+efficient+neural+conversion+of+human+ES+and+iPS+cells+by+dual+inhibition+of+SMAD+signaling.">Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.</a> Nature Biotechnology. 27 (3), 275-280 (2009).</li><li>Takasato, 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=Directing+human+embryonic+stem+cell+differentiation+towards+a+renal+lineage+generates+a+self-organizing+kidney.">Directing human embryonic stem cell differentiation towards a renal lineage generates a self-organizing kidney.</a> Nature cell biology. 16 (1), 118-126 (2014).</li><li>Zimmer, B., 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=Evaluation+of+Developmental+Toxicants+and+Signaling+Pathways+in+a+Functional+Test+Based+on+the+Migration+of+Human+Neural+Crest+Cells.">Evaluation of Developmental Toxicants and Signaling Pathways in a Functional Test Based on the Migration of Human Neural Crest Cells.</a> Environmental Health Perspectives. 120 (8), 1116-1122 (2012).</li><li>Bosman, 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=Molecular+and+Functional+Evidence+of+HCN4+and+Caveolin-3+Interaction+During+Cardiomyocyte+Differentiation+from+Human+Embryonic+Stem+Cells.">Molecular and Functional Evidence of HCN4 and Caveolin-3 Interaction During Cardiomyocyte Differentiation from Human Embryonic Stem Cells.</a> Stem Cells and Development. 22 (11), 1717-1727 (2013).</li><li>Sartiani, 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=Developmental+changes+in+cardiomyocytes+differentiated+from+human+embryonic+stem+cells:+A+molecular+and+electrophysiological+approach.">Developmental changes in cardiomyocytes differentiated from human embryonic stem cells: A molecular and electrophysiological approach.</a> Stem Cells. 25 (5), 1136-1144 (2007).</li><li>Xu, X. Q., 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=Chemically+defined+medium+supporting+cardiomyocyte+differentiation+of+human+embryonic+stem+cells.">Chemically defined medium supporting cardiomyocyte differentiation of human embryonic stem cells.</a> Differentiation. 76 (9), 958-970 (2008).</li><li>Pal, R., Mamidi, M. K., Das, A. K., Bhonde, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+analysis+of+cardiomyocyte+differentiation+from+human+embryonic+stem+cells+under+3-D+and+2-D+culture+conditions.">Comparative analysis of cardiomyocyte differentiation from human embryonic stem cells under 3-D and 2-D culture conditions.</a> Journal of Bioscience and Bioengineering. 115 (2), 200-206 (2013).</li><li>Subramanian, 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=Spheroid+Culture+for+Enhanced+Differentiation+of+Human+Embryonic+Stem+Cells+to+Hepatocyte-Like+Cells.">Spheroid Culture for Enhanced Differentiation of Human Embryonic Stem Cells to Hepatocyte-Like Cells.</a> Stem Cells and Development. 23 (2), 124-131 (2014).</li><li>Sivertsson, L., Synnergren, J., Jensen, J., Bjorquist, P., Ingelman-Sundberg, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+Differentiation+and+Maturation+of+Human+Embryonic+Stem+Cells+Cultured+in+a+Perfused+Three-Dimensional+Bioreactor.">Hepatic Differentiation and Maturation of Human Embryonic Stem Cells Cultured in a Perfused Three-Dimensional Bioreactor.</a> Stem Cells and Development. 22 (4), 581-594 (2013).</li><li>Jagtap, 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=Cytosine+arabinoside+induces+ectoderm+and+inhibits+mesoderm+expression+in+human+embryonic+stem+cells+during+multilineage+differentiation.">Cytosine arabinoside induces ectoderm and inhibits mesoderm expression in human embryonic stem cells during multilineage differentiation.</a> British Journal of Pharmacology. 162 (8), 1743-1756 (2011).</li><li>Krug, 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=Human+embryonic+stem+cell-derived+test+systems+for+developmental+neurotoxicity:+a+transcriptomics+approach.">Human embryonic stem cell-derived test systems for developmental neurotoxicity: a transcriptomics approach.</a> Archives of Toxicology. 87 (1), 123-143 (2013).</li><li>Leist, 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=Test+systems+of+developmental+toxicity:+state-of-the+art+and+future+perspectives.">Test systems of developmental toxicity: state-of-the art and future perspectives.</a> Archives of Toxicology. 87 (12), 2037-2042 (2013).</li><li>Itskovitz-Eldor, 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=Differentiation+of+human+embryonic+stem+cells+into+embryoid+bodies+compromising+the+three+embryonic+germ+layers.">Differentiation of human embryonic stem cells into embryoid bodies compromising the three embryonic germ layers.</a> Molecular medicine. 6 (2), 88-95 (2000).</li><li>Khoo, M. L. 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=Growth+and+differentiation+of+embryoid+bodies+derived+from+human+embryonic+stem+cells:+Effect+of+glucose+and+basic+fibroblast+growth+factor.">Growth and differentiation of embryoid bodies derived from human embryonic stem cells: Effect of glucose and basic fibroblast growth factor.</a> Biology of Reproduction. 73 (6), 1147-1156 (2005).</li><li>Son, M. Y., Kim, H. J., Kim, M. J., Cho, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physical+Passaging+of+Embryoid+Bodies+Generated+from+Human+Pluripotent+Stem+Cells.">Physical Passaging of Embryoid Bodies Generated from Human Pluripotent Stem Cells.</a> Plos One. 6 (5), (2011).</li><li>Winkler, J., Sotiriadou, I., Chen, S., Hescheler, J., Sachinidis, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+potential+of+embryonic+stem+cells+combined+with+-omics+technologies+as+model+systems+for+toxicology.">The potential of embryonic stem cells combined with -omics technologies as model systems for toxicology.</a> Current medicinal chemistry. 16 (36), 4814-4827 (2009).</li><li>Gunaseeli, I., Doss, M. X., Antzelevitch, C., Hescheler, J., Sachinidis, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induced+pluripotent+stem+cells+as+a+model+for+accelerated+patient-+and+disease-specific+drug+discovery.">Induced pluripotent stem cells as a model for accelerated patient- and disease-specific drug discovery.</a> Current medicinal chemistry. 17 (8), 759-766 (2010).</li><li>Miller, M. T., Stromland, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Teratogen+update:+Thalidomide:+A+review,+with+a+focus+on+ocular+findings+and+new+potential+uses.">Teratogen update: Thalidomide: A review, with a focus on ocular findings and new potential uses.</a> Teratology. 60 (5), 306-321 (1999).</li><li>Newman, C. G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Teratogen+Update+-+Clinical+Aspects+of+Thalidomide+Embryopathy+-+A+Continuing+Preoccupation.">Teratogen Update - Clinical Aspects of Thalidomide Embryopathy - A Continuing Preoccupation.</a> Teratology. 32 (1), 133-144 (1985).</li><li>Stern, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+assessment+of+the+teratogenic+potential+of+drugs+in+humans.">In vivo assessment of the teratogenic potential of drugs in humans.</a> Obstetrics and gynecology. 58 (5 Suppl), 3S-8S (1981).</li><li>Lynch, H. T., Reich, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diethylstilbestrol,+Genetics,+Teratogenesis,+and+Tumor+Spectrum+in+Humans.">Diethylstilbestrol, Genetics, Teratogenesis, and Tumor Spectrum in Humans.</a> Medical Hypotheses. 16 (3), 315-332 (1985).</li><li>Satoh, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+teratology+of+mercury+and+its+compounds.">Behavioral teratology of mercury and its compounds.</a> Tohoku Journal of Experimental Medicine. 201 (1), 1-9 (2003).</li><li>Meganathan, 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=Identification+of+Thalidomide-Specific+Transcriptomics+and+Proteomics+Signatures+during+Differentiation+of+Human+Embryonic+Stem+Cells.">Identification of Thalidomide-Specific Transcriptomics and Proteomics Signatures during Differentiation of Human Embryonic Stem Cells.</a> Plos One. 7 (8), (2012).</li><li>Balmer, N. V., 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=Epigenetic+changes+and+disturbed+neural+development+in+a+human+embryonic+stem+cell-based+model+relating+to+the+fetal+valproate+syndrome.">Epigenetic changes and disturbed neural development in a human embryonic stem cell-based model relating to the fetal valproate syndrome.</a> Human Molecular Genetics. 21 (18), 4104-4114 (2012).</li><li>Smirnova, L., Hogberg, H. T., Leist, M., Hartung, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+neurotoxicity+-+Challenges+in+the+21st+Century+and+In+Vitro+Opportunities.">Developmental neurotoxicity - Challenges in the 21st Century and In Vitro Opportunities.</a> ALTEX. 31 (2), 129-156 (2014).</li><li>Leist, M., Efremova, L., Karreman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Food+for+thought+...+considerations+and+guidelines+for+basic+test+method+descriptions+in+toxicology.">Food for thought ... considerations and guidelines for basic test method descriptions in toxicology.</a> ALTEX. 27 (4), 309-317 (2010).</li><li>Zimmer, B., 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=Coordinated+waves+of+gene+expression+during+neuronal+differentiation+of+embryonic+stem+cells+as+basis+for+novel+approaches+to+developmental+neurotoxicity+testing.">Coordinated waves of gene expression during neuronal differentiation of embryonic stem cells as basis for novel approaches to developmental neurotoxicity testing.</a> Cell Death and Differentiation. 18 (3), 383-395 (2011).</li><li>Kraljevic, S., Stambrook, P. J., Pavelic, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerating+drug+discovery.">Accelerating drug discovery.</a> EMBO Reports. 5 (9), 837-842 (2004).</li></ol>

The authors have nothing to disclose.

传统方法毒理试验涉及广泛的动物研究中从而使测试昂贵和费时。此外，由于种间差异的临床前动物安全性研究并不总是有效的预测与人类有关的潜在药物毒性作用。虽然非人类灵长类动物是最可预测的，还是强烈的伦理和socioeconomical需求正在迅速被现代社会提高发展中国家在体外试验敏感和强大 有关人类的安全系统。 人类胚胎干细胞的独特能力分化成所有的?...

人类胚胎干细胞（hESC细胞）的分化成各种细胞的能力打开了体外毒性试验1中 ，疾病建模和再生医学2的新时代。干细胞被赋予的能力，自我复制，以保持其多能状态，并分化成特定细胞3,4。的人类胚胎干细胞的特性（容量来区分所有主要的细胞类型），也发现在其它人多能干细胞，如人类诱导多能干细胞（hiPSC）或通过核移植5产生的细胞。例如，许多不同的hESC细胞系已分化为神经元6，肾细胞7，神经嵴细胞8，心肌9-12，或肝细胞样细胞13,14。此外，人类胚胎干细胞可自发地分化成胚状体（EB）19,20所有三种胚层15-18的细胞。 Ë阿尔利胚胎发育是通过与已捕获在mRNA水平通过使用微阵列技术15转录的不同胚层的多种基因的差异表达调控。这些努力使建立基于人类胚胎干细胞/ hiPSC和转录组分析（综述见21,22）器官特异性毒性模型。这些车型有优势的传统使用实验动物的毒理学研究，如使用实验动物并不总是预测人类安全的临床前研究。在患者中遇到的药物诱导的毒性往往与人类和实验动物之间是不同的代谢或信号传导过程。的物种差异防止了可靠的早期检测发育毒性在人类，例如药物如沙利度胺23,24和己烯雌酚25,26从市场上撤回由于致畸。塔利domide并没有表现出在大鼠或小鼠任何发育毒性。环境化学品如甲基汞27导致相对于神经系统中的各种物种产前发育毒性，但人的表现已经很难在动物模型。为了解决种属特异性问题的问题，科学家们根据不同的项目工作的基础上的干细胞一样重新保护，ESNATS，侦探等所从事的不同型号的发展胚胎毒性，神经毒性，心脏毒性，肝毒性和肾毒性涉嫌使用对人体有毒物质影响人类。根据欧洲联盟项目的胚胎干细胞为基础的新型替代测试策略（ESNATS）"五个测试系统已经建立。一个测试系统的所谓UKK（UniversitätsķlinikumķOLN）测试系统部分捕捉人类早期胚胎发育。在这个System人类胚胎H9细胞分化中15和胚层特定的签名已被抓获转录使用Affymetrix芯片平台简介三个胚层（外胚层，内胚层和中胚层）。像沙利度胺28，丙戊酸，甲基汞16,17，或阿糖胞苷15各个发育毒物在该系统已经过测试，并且具体毒物基因签名已被获得。在第二测试系统，即所谓的UKN1（U。高校ķonsta N个Z的）测试系统1中，H9细胞分化到神经外胚层祖细胞（NEP）6天。这是通过高表达的神经基因标记如PAX6和OTX2证实。在分化6天，NEP细胞已经暴露于发育神经毒物如丙戊酸钠，甲基汞。毒物特异性解除管制的转录概况已obtai通过使用Affymetrix芯片平台定义16,29以及。 对于21 世纪的毒理学的新远景设想，测试系统不仅产生的表型的描述，如组织病理学体内 ，或转录变化在长期毒物孵化结束。它，而表明试验提供机械信息3，而该信息可以被映射到所谓的不利后果的途径（AOP），提供了一个科学原理为危险影响30。为了提供这种信息，应用的测试系统必须高度控制质量31，作为例如通过稳健标准操作程序记载。此外，依赖于时间的变化需要被映射为高的分辨率。这就要求测试系统同步变化32。这里所描述的UKN1和UKK测试系统进行了优化这些需求。

<table><tbody><tr><td>DMEM/F-12</td><td>Life Technologies</td><td>11320082</td><td>Dulbecco&#039;s Modified Eagle Medium:Nutrient Mixture F-12</td></tr><tr><td>KOSR</td><td>Life Technologies</td><td>10828028</td><td>Knockout Serum Replacement</td></tr><tr><td>GlutaMAX</td><td>Life Technologies</td><td>35050061</td><td>GlutaMAX supplement</td></tr><tr><td>NEAA</td><td>Life Technologies</td><td>11140050</td><td>MEM Nonessential Amino Acids Solution</td></tr><tr><td>DPBS</td><td>Life Technologies</td><td>14190-0144</td><td>Dulbecco&#039;s Phosphate-Buffered Saline, without calcium, without magnesium</td></tr><tr><td>mTeSR medium</td><td>Stemcell Technologies</td><td>5850</td><td></td></tr><tr><td>Pluronic F-127</td><td>Sigma</td><td>P2443-250G</td><td></td></tr><tr><td>V bottom plate</td><td>VWR</td><td>734-0483</td><td>Plate,Microwell,V BTTM,96 Well,Sterile 1 * 50 ST</td></tr><tr><td>V bottom plate lid</td><td>VWR</td><td>634-0011</td><td>Lid, Microtitre plates, Cond. Ring 1 * 50 ST</td></tr><tr><td>Pen/Strep</td><td>Life Technologies</td><td>15140-122</td><td>Penicillin-Streptomycin, Liquid</td></tr><tr><td>Distilled Water</td><td>Life Technologies</td><td>15230-089.</td><td>Sterile Distilled Water</td></tr><tr><td>Human FGF-2 (bFGF)</td><td>Millipore</td><td>GF003AF-100UG</td><td>Fibroblast Growth Factor basic, human recombinant, animal-free</td></tr><tr><td>Filter 0.22 &amp;mu;m</td><td>Millipore</td><td>SCGPU02RE</td><td>Stericup-GP, 0.22 &amp;mu;m, polyethersulfone, 250 ml, radio-sterilized</td></tr><tr><td>StemPro EZPassage&lt;sup&gt;TM&lt;/sup&gt; Disposablte</td><td>Invitrogen</td><td>23181010</td><td></td></tr><tr><td>BD MatrigelTM, hESC qualified Matrix</td><td>Stemcell Technologies</td><td>354277</td><td>5 ml vial</td></tr><tr><td>DMSO</td><td>Sigma</td><td>D-2650</td><td></td></tr><tr><td>RNAlater Stabilization Solution</td><td>Life Technologies</td><td>AM7020</td><td>It stabilizes and protect the RNA integrity in unfrozen samples.</td></tr><tr><td>70 &amp;mu;m Cell Strainer</td><td>Becton Dickinson</td><td>352350</td><td>Cell strainer with 70 &amp;mu;m Nylon mesh</td></tr><tr><td>35 &amp;mu;m Lid cell strainer, 5&amp;nbsp;ml tube</td><td>Becton Dickinson</td><td>352235</td><td>5 ml polystyrene round bottom test tube, with a cell strainer cap (35&amp;nbsp;&amp;mu;m)</td></tr><tr><td>50 ml sterile Polypropylene tube</td><td>Greiner Bio-One</td><td>227261</td><td>50 ml Polypropylene tube with conical bottom, Sterile</td></tr><tr><td>T75 flask</td><td>Greiner Bio-One</td><td>658175</td><td>CELLSTAR Filter Cap Cell Culture 75 cm&lt;sup&gt;2&lt;/sup&gt; Flasks</td></tr><tr><td>TRIzol</td><td>Life Technologies</td><td>10296010</td><td></td></tr><tr><td>96 well optical bottom plates</td><td>Thermo Scientific</td><td>165305</td><td></td></tr><tr><td>CellTiter-Blue</td><td>Promega</td><td>G8081</td><td></td></tr><tr><td>Accutase</td><td>PAA</td><td>L11-007</td><td></td></tr><tr><td>Apotransferin</td><td>Sigma-Aldrich</td><td>T-2036</td><td></td></tr><tr><td>Dispase</td><td>Worthington Biochemicals</td><td>LS002104</td><td></td></tr><tr><td>Dorsomorphin</td><td>Tocris Bioscience</td><td>3093</td><td></td></tr><tr><td>EDTA</td><td>Roth</td><td>8043.2</td><td></td></tr><tr><td>FBS</td><td>PAA</td><td>A15-101</td><td></td></tr><tr><td>FGF-2</td><td>R&amp;amp;D Systems</td><td>233-FB</td><td></td></tr><tr><td>Gelatine</td><td>Sigma-Aldrich</td><td>G1890-100G</td><td></td></tr><tr><td>Glucose</td><td>Sigma-Aldrich</td><td>G7021-100G</td><td></td></tr><tr><td>GlutaMAX</td><td>Gibco Invitrogen</td><td>35050-038</td><td></td></tr><tr><td>HEPES</td><td>Gibco Invitrogen</td><td>15630-056</td><td></td></tr><tr><td>Insulin</td><td>Sigma-Aldrich</td><td>I-6634</td><td></td></tr><tr><td>Knockout DMEM</td><td>Gibco Invitrogen</td><td>10829-018</td><td></td></tr><tr><td>Matrigel</td><td>BD Biosciences</td><td>354234</td><td></td></tr><tr><td>Noggin</td><td>R&amp;amp;D Systems</td><td>719-NG</td><td></td></tr><tr><td>PBS</td><td>Biochrom AG</td><td>L1825</td><td></td></tr><tr><td>Progesteron</td><td>Sigma-Aldrich</td><td>P7556</td><td></td></tr><tr><td>Putrescine</td><td>Sigma-Aldrich</td><td>P-5780</td><td></td></tr><tr><td>ROCK inhibitor Y-27632</td><td>Tocris Biosciences</td><td>1254</td><td></td></tr><tr><td>SB431542</td><td>Tocris Biosciences</td><td>1614</td><td></td></tr><tr><td>SDS</td><td>Bio-Rad</td><td>161-0416</td><td></td></tr><tr><td>Selenium</td><td>Sigma-Aldrich</td><td>S-5261</td><td></td></tr><tr><td>&amp;beta;-Mercaptoethanol</td><td>Gibco Invitrogen</td><td>31350-010</td><td></td></tr><tr><td colspan="4">[header]</td></tr><tr><td colspan="4">&lt;strong&gt;List of Kits&lt;/strong&gt;</td></tr><tr><td>RNeasy Mini Kit (250)</td><td>QIAGEN</td><td>74106</td><td></td></tr><tr><td>GeneChip Hybridization, Wash, and Stain Kit</td><td>Affymetrix</td><td>900721, 22, 23</td><td>This kit provides all reagents required for hybridization wash and staining of microarrays.</td></tr><tr><td>Rnase-Free DNase Set</td><td>QIAGEN</td><td>79254</td><td></td></tr><tr><td colspan="4">[header]</td></tr><tr><td colspan="4">&lt;strong&gt;List of equipment&lt;/strong&gt;</td></tr><tr><td>Inverted microscope</td><td>Olympus</td><td></td><td>IX71</td></tr><tr><td>Genechip Hybridisation Oven - 645</td><td>Affymetrix</td><td></td><td></td></tr><tr><td>Genechip Fluidics Station-450</td><td>Affymetrix</td><td></td><td></td></tr><tr><td>Affymetrix Gene-Chip Scanner-3000-7 G</td><td>Affymetrix</td><td></td><td></td></tr><tr><td>Spectramax M5</td><td>Molecular Devices</td><td></td><td>&lt;br /&gt;&amp;#160;&lt;br /&gt;&amp;#160;&lt;br /&gt;&amp;#160;</td></tr><tr><td colspan="4">[header]</td></tr><tr><td colspan="4">&lt;strong&gt;List of softwares&lt;/strong&gt;</td></tr><tr><td>Prism 4</td><td></td><td></td><td></td></tr><tr><td>Affymetrix GCOS</td><td></td><td></td><td></td></tr><tr><td>Partek Genomic Suite 6.25</td><td></td><td></td><td></td></tr><tr><td>Online tools for Functional annotation&lt;br/&gt; DAVID&lt;br/&gt; Onto-tools Intelligent Systems and Bioinformatics Laboratory</td><td></td><td></td><td></td></tr></tbody></table>

human pluripotent stem cell based developmental toxicity assays for chemical safety screening and systems biology data generation

高效的协议来区分人多能干细胞，以组合各种组织与组学技术开辟了新的视野为潜在药物的体外毒性测试。为了提供这种试验坚实的科学基础，这将是非常重要的获得发展的时间进程和底层的监管机制，系统生物学方法的定量信息。两个实验都因此被调整为在这里这些要求。在UKK测试系统中，人胚胎干细胞（hESC细胞）（或其它多能细胞）都留给自发分化为胚状体14天，以允许产生所有三个胚层的细胞。该系统概括的人类早期胚胎发育的关键步骤，并且它可以预测人特异性早期胚胎毒性/致畸性，如果细胞分化过程中暴露于化学品。所述UKN1测试系统是基于人类胚胎干细胞分化为一个populat神经外胚层祖离子（NEP）细胞6天。该系统概括早期神经发育，并预测早期发育神经毒性和化学品引发的表观遗传变化。两个系统，在与转录微阵列研究结合，适合于识别的毒性的生物标志物。此外，它们也可以组合使用以产生用于系统生物学分析的输入数据。这些测试系统具有比传统的毒理学研究需要大量的动物的优点。该测试系统可能有助于减少药物开发和化学安全性评价的费用。他们的组合揭示尤其是对可能影响具体化合物的神​​经发育光。

在UKK测试系统甲基汞暴露与H9的EB进行的细胞毒性测定，得到的IC 10值（10％减少活力的）甲基汞的细胞毒性（ 图1）。我们还进行了微阵列基础（Affymetrix公司的平台）的生物标志物研究。的H9的EB已暴露于甲基汞（0.25和1μM）14天。在第14天，将样品已收集使用Trizol和分离RNA。使用人类基因组U133加2.0阵列芯片进行转录谱。的数据进行了分析以将Partek基因组套?...

Watch this Scientific Journal Video about Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation at JoVE.com

Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation

该协议描述了基于人胚胎干细胞和转录研究2 在体外发育毒性试验系统（UKK和UKN1）。测试系统预测人类发育毒性危险，并可能有助于减少动物研究中，成本和所需的化学安全测试的时间。

基于发育毒性测定化学品安全筛选和系统生物学数据生成人类多能干细胞

Efficient protocols to differentiate human pluripotent stem cells to various tissues in combination with -omics technologies opened up new horizons for in ...

human-pluripotent-stem-cell-based-developmental-toxicity-assays-for

Research

JoVE Journal

Developmental Biology

12.5K 次观看.  University of Cologne. 该协议描述了基于人胚胎干细胞和转录研究2 在体外发育毒性试验系统（UKK和UKN1）。测试系统预测人类发育毒性危险，并可能有助于减少动物研究中，成本和所需的化学安全测试的时间。 

视频: Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation

Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing

Human pluripotent stem cells can differentiate into various cell types that can be applied to human-based in vitro toxicity assays. One major advantage is that the reprogramming of somatic cells to produce human induced pluripotent stem cells (hiPSCs) avoids the ethical and legislative issues related to the use of human embryonic stem cells (hESCs). HiPSCs can be expanded and efficiently differentiated into different types of neuronal and glial cells, serving as test systems for toxicity testing and, in particular, for the assessment of different pathways involved in neurotoxicity. This work describes a protocol for the differentiation of hiPSCs into mixed cultures of neuronal and glial cells. The signaling pathways that are regulated and/or activated by neuronal differentiation are defined. This information is critical to the application of the cell model to the new toxicity testing paradigm, in which chemicals are assessed based on their ability to perturb biological pathways. As a proof of concept, rotenone, an inhibitor of mitochondrial respiratory complex I, was used to assess the activation of the Nrf2 signaling pathway, a key regulator of the antioxidant-response-element-(ARE)-driven cellular defense mechanism against oxidative stress.

Human induced pluripotent stem cells (hiPSCs) are considered a powerful tool for drug and chemical screening and for the development of new in vitro models for toxicity testing, including neurotoxicity. Here, a detailed protocol for the differentiation of hiPSCs into neurons and glia is described.

将人诱导的多能干细胞分化为神经元和神经胶质的混合培养物进行神经毒性测试的方案

Human pluripotent stem cells can differentiate into various cell types that can be applied to human-based in vitro toxicity assays. One major advantage is ...

科研

发育生物学

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and maintenance issues. On the contrary, the nematode Caenorhabditis elegans (C. elegans) has advantages to make it an ideal choice for toxicity testing: a short lifespan, easy cultivation, and efficient reproduction. Here, we describe a protocol for the automatic phenotypic profiling of C. elegans in a 384-well plate. The nematode worms are cultured in a 384-well plate with liquid medium and chemical treatment, and videos are taken of each well to quantify the chemical influence on 33 worm features. Experimental results demonstrate that the quantified phenotype features can classify and predict the acute toxicity for different chemical compounds and establish a priority list for further traditional chemical toxicity assessment tests in a rodent model.

A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans

A quantitative method has been developed to identify and predict the acute toxicity of chemicals by automatically analyzing the phenotypic profiling of Caenorhabditis elegans. This protocol describes how to treat worms with chemicals in a 384-well plate, capture videos, and quantify toxicological related phenotypes.

一种用自动表型分析预测线虫化学毒性的高通量测定

Applying toxicity testing of chemicals in higher order organisms, such as mice or rats, is time-consuming and expensive, due to their long lifespan and ...

环境科学

Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery

The ability to differentiate human induced pluripotent stem cells (iPSCs) into hepatocyte-like cells (HLCs) provides new opportunities to study inborn errors in hepatic metabolism. However, to provide a platform that supports the identification of small molecules that can potentially be used to treat liver disease, the procedure requires a culture format that is compatible with screening thousands of compounds. Here, we describe a protocol using completely defined culture conditions, which allow the reproducible differentiation of human iPSCs to hepatocyte-like cells in 96-well tissue culture plates. We also provide an example of using the platform to screen compounds for their ability to lower Apolipoprotein B (APOB) produced from iPSC-derived hepatocytes generated from a familial hypercholesterolemia patient. The availability of a platform that is compatible with drug discovery should allow researchers to identify novel therapeutics for diseases that affect the liver.

The protocol presented here describes a platform for identifying small molecules for the treatment of liver disease. A step-by-step description is presented detailing how to differentiate iPSCs into cells with hepatocyte characteristics in 96-well plates, and to use the cells to screen for small molecules with potential therapeutic activity.

人诱导多潜能干细胞衍生肝细胞的药物发现方法

The ability to differentiate human induced pluripotent stem cells (iPSCs) into hepatocyte-like cells (HLCs) provides new opportunities to study inborn ...

Semi-automated Production of Hepatocyte Like Cells from Pluripotent Stem Cells

Human pluripotent stem cells represent a renewable source of human tissue. Our research is focused on generating human liver tissue from human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). Current differentiation procedures generate human hepatocyte-like cells (HLCs) displaying a mixture of fetal and adult traits. To improve cell phenotype, we have fully defined our differentiation procedure and the cell niche, resulting in the generation of cell populations which display improved gene expression and&#160;function. While these studies mark progress, the ability to generate large quantities of multi well plates for screening has been limited by labour intensive procedures and batch to batch variation. To tackle this issue, we have developed a semi-automated platform to differentiate pluripotent stem cells into HLCs. Stem cell seeding and differentiation were performed using liquid handling and automatic pipetting systems in&#160;96-well plate format. Following the differentiation, cell phenotype was analyzed using automated microscopy and a multi well luminometer.

This protocol describes a semi-automated approach to produce hepatocyte-like cells from human pluripotent stem cells in a 96 well plate format. This process is rapid and cost-effective, allowing the production of quality assured batches of hepatocyte-like cells for basic and applied human research.

多能干细胞半自动化生产肝样细胞的研究

Human pluripotent stem cells represent a renewable source of human tissue. Our research is focused on generating human liver tissue from human embryonic ...

A Neurite Outgrowth Assay and Neurotoxicity Assessment with Human Neural Progenitor Cell-Derived Neurons

Neurite outgrowth assay and neurotoxicity assessment are two major studies that can be performed using the presented method herein. This protocol provides reliable analysis of neuronal morphology together with quantitative measurements of modifications on neurite length and synaptic protein localization and abundance upon treatment with small molecule compounds. In addition to the application of the presented method in neurite outgrowth studies, neurotoxicity assessment can be performed to assess, distinguish and rank commercial chemical compounds based on their potential developmental neurotoxicity effect.
Even though cell lines are nowadays widely used in compound screening assays in neuroscience, they often differ genetically and phenotypically from their tissue origin. Primary cells, on the other hand, maintain important markers and functions observed in vivo. Therefore, due to the translation potential and physiological relevance that these cells could offer neurite outgrowth assay and neurotoxicity assessment can considerably benefit from using human neural progenitor cells (hNPCs) as the primary human cell model.
The presented method herein can be utilized to screen for the ability of compounds to induce neurite outgrowth and neurotoxicity by taking advantage of the human neural progenitor cell-derived neurons, a cell model closely representing human biology.&#34;

The presented protocol describes a method for a neurite outgrowth assay and neurotoxicity assessment of small molecule compounds.

神经石生长分析与神经毒性评估与人类神经原生细胞衍生神经元

Neurite outgrowth assay and neurotoxicity assessment are two major studies that can be performed using the presented method herein. This protocol provides ...

Robust Generation of Hepatocyte-like Cells from Human Embryonic Stem Cell Populations

Despite progress in modelling human drug toxicity, many compounds fail during clinical trials due to unpredicted side effects. The cost of clinical studies are substantial, therefore it is essential that more predictive toxicology screens are developed and deployed early on in drug development (Greenhough et al 2010). Human hepatocytes represent the current gold standard model for evaluating drug toxicity, but are a limited resource that exhibit variable function. Therefore, the use of immortalised cell lines and animal tissue models are routinely employed due to their abundance. While both sources are informative, they are limited by poor function, species variability and/or instability in culture (Dalgetty et al 2009). Pluripotent stem cells (PSCs) are an attractive alternative source of human hepatocyte like cells (HLCs) (Medine et al 2010). PSCs are capable of self renewal and differentiation to all somatic cell types found in the adult and thereby represent a potentially inexhaustible source of differentiated cells. We have developed a procedure that is simple, highly efficient, amenable to automation and yields functional human HLCs (Hay et al 2008 ; Fletcher et al 2008 ; Hannoun et al 2010 ; Payne et al 2011 and Hay et al 2011). We believe our technology will lead to the scalable production of HLCs for drug discovery, disease modeling, the construction of extra-corporeal devices and possibly cell based transplantation therapies.

This article will focus on the generation of human hepatic endoderm from human embryonic stem cell populations.

肝细胞样细胞从人类胚胎干细胞群的鲁棒代

Despite progress in modelling human drug toxicity, many compounds fail during clinical trials due to unpredicted side effects. The cost of clinical ...

生物学

Toxicity Screens in Human Retinal Organoids for Pharmaceutical Discovery

Organoids provide a promising platform to study disease mechanism and treatments, directly in the context of human tissue with the versatility and throughput of cell culture. Mature human retinal organoids are utilized to screen potential pharmaceutical treatments for the age-related retinal degenerative disease macular telangiectasia type 2 (MacTel).
We have recently shown that MacTel can be caused by elevated levels of an atypical lipid species, deoxysphingolipids (deoxySLs). These lipids are toxic to the retina and may drive the photoreceptor loss that occurs in MacTel patients. To screen drugs for their ability to prevent deoxySL photoreceptor toxicity, we generated human retinal organoids from a non-MacTel induced pluripotent stem cell (iPSC) line and matured them to a post-mitotic age where they develop all of the neuronal lineage-derived cells of the retina, including functionally mature photoreceptors. The retinal organoids were treated with a deoxySL metabolite and apoptosis was measured within the photoreceptor layer using immunohistochemistry. Using this toxicity model, pharmacological compounds that prevent deoxySL-induced photoreceptor death were screened. Using a targeted candidate approach, we determined that fenofibrate, a drug commonly prescribed for the treatment of high cholesterol and triglycerides, can also prevent deoxySL toxicity in the cells of the retina.
The toxicity screen successfully identified an FDA-approved drug that can prevent photoreceptor death. This is a directly actionable finding owing to the highly disease-relevant model tested. This platform can be easily modified to test any number of metabolic stressors and potential pharmacological interventions for future treatment discovery in retinal diseases.

Here we present a step-by-step protocol to generate mature human retinal organoids and utilize them in a photoreceptor toxicity assay to identify pharmaceutical candidates for the age-related retinal degenerative disease macular telangiectasia type 2 (MacTel).

用于药物发现的人视网膜类器官毒性筛选

Organoids provide a promising platform to study disease mechanism and treatments, directly in the context of human tissue with the versatility and ...

生物工程

Hybrid Cell Analysis System to Assess Structural and Contractile Changes of Human iPSC-Derived Cardiomyocytes for Preclinical Cardiac Risk Evaluation

Cardiac contractility assessment is of immense importance for the development of new therapeutics and their safe transition into clinical stages. While human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold promise to serve as a human-relevant model in preclinical phases of drug discovery and safety pharmacology, their maturity is still controversial in the scientific community and under constant development. We present a hybrid contractility and impedance/extracellular field potential (EFP) technology, adding significant pro-maturation features to an industry-standard 96-well platform.
The impedance/EFP system monitors cellular functionality in real-time. Besides the beat rate of contractile cells, the electrical impedance spectroscopy readouts detect compound-induced morphological changes like cell density and integrity of the cellular monolayer. In the other component of the hybrid cell analysis system, the cells are cultured on bio-compliant membranes that mimic the mechanical environment of real heart tissue. This physiological environment supports the maturation of hiPSC-CMs in vitro, leading to more adult-like contractile responses including positive inotropic effects after treatment with isoproterenol, S-Bay K8644, or omecamtiv mecarbil. Parameters such as the amplitude of contraction force (mN/mm2) and beat duration also reveal downstream effects of compounds with influence on electrophysiological properties and calcium handling.
The hybrid system provides the ideal tool for holistic cell analysis, allowing preclinical cardiac risk assessment beyond the current perspectives of human-relevant cell-based assays.

The analysis of changes in contractile function and cellular integrity of human iPSC-derived cardiomyocytes is of immense importance for nonclinical drug development. A hybrid 96-well cell analysis system addresses both parameters in a real-time and physiological manner for reliable, human-relevant results, necessary for a safe transition into clinical stages.

用于评估人iPSC来源心肌细胞结构和收缩变化的杂交细胞分析系统，用于临床前心脏风险评估

Cardiac contractility assessment is of immense importance for the development of new therapeutics and their safe transition into clinical stages. While ...

High-Throughput Cardiotoxicity Screening Using Mature Human Induced Pluripotent Stem Cell-Derived Cardiomyocyte Monolayers

Human induced stem cell-derived cardiomyocytes (hiPSC-CMs) are used to replace and reduce the dependence on animals and animal cells for preclinical cardiotoxicity testing. In two-dimensional monolayer formats, hiPSC-CMs recapitulate the structure and function of the adult human heart muscle cells when cultured on an optimal extracellular matrix (ECM). A human perinatal stem cell-derived ECM (maturation-inducing extracellular matrix-MECM) matures the hiPSC-CM structure, function, and metabolic state in 7 days after plating. 
Mature hiPSC-CM monolayers also respond as expected to clinically relevant medications, with a known risk of causing arrhythmias and cardiotoxicity. The maturation of hiPSC-CM monolayers was an obstacle to the widespread adoption of these valuable cells for regulatory science and safety screening, until now. This article presents validated methods for the plating, maturation, and high-throughput, functional phenotyping of hiPSC-CM electrophysiological and contractile function. These methods apply to commercially available purified cardiomyocytes, as well as stem cell-derived cardiomyocytes generated in-house using highly efficient, chamber-specific differentiation protocols. 
High-throughput electrophysiological function is measured using either voltage-sensitive dyes (VSDs; emission: 488 nm), calcium-sensitive fluorophores (CSFs), or genetically encoded calcium sensors (GCaMP6). A high-throughput optical mapping device is used for optical recordings of each functional parameter, and custom dedicated software is used for electrophysiological data analysis. MECM protocols are applied for medication screening using a positive inotrope (isoprenaline) and human Ether-a-go-go-related gene (hERG) channel-specific blockers. These resources will enable other investigators to successfully utilize mature hiPSC-CMs for high-throughput, preclinical cardiotoxicity screening, cardiac medication efficacy testing, and cardiovascular research.

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offer an alternative to using animals for preclinical cardiotoxicity screening. A limitation to the widespread adoption of hiPSC-CMs in preclinical toxicity screening is the immature, fetal-like phenotype of the cells. Presented here are protocols for robust and rapid maturation of hiPSC-CMs.

使用成熟的人诱导多能干细胞来源心肌细胞单层进行高通量心脏毒性筛选

Human induced stem cell-derived cardiomyocytes (hiPSC-CMs) are used to replace and reduce the dependence on animals and animal cells for preclinical ...

High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents (HPHC)

Cigarette smoke (CS) is a major risk factor for cardiovascular and lung diseases. Because CS is a complex aerosol containing more than 7,000 chemicals1 it is challenging to assess the contributions of individual constituents to its overall toxicity. Toxicological profiles of individual constituents as well as mixtures can be however established in vitro, by applying high through-put screening tools, which enable the profiling of Harmful and Potentially Harmful Constituents (HPHCs) of tobacco smoke, as defined by the U.S. Food and Drug Administration (FDA).2
For an initial assessment, an impedance-based instrument was used for a real-time, label-free assessment of the compound&#39;s toxicity. The instrument readout relies on cell adhesion, viability and morphology that all together provide an overview of the cell status. A dimensionless parameter, named cell index, is used for quantification. A set of different staining protocols was developed for a fluorescence imaging-based investigation and a HCS platform was used to gain more in-depth information on the kind of cytotoxicity elicited by each HPHC.
Of the 15 constituents tested, only five were selected for HCS-based analysis as they registered a computable LD50 (&#60; 20 mM). These included 1-aminonaphtalene, Arsenic (V), Chromium (VI), Crotonaldehyde and Phenol. Based on their effect in the HCS, 1-aminonaphtalene and Phenol could be identified to induce mitochondrial dysfunction, and, together with Chromium (VI) as genotoxic based on the increased histone H2AX phosphorylation. Crotonaldehyde was identified as an oxidative stress inducer and Arsenic as a stress kinase pathway activator.
This study demonstrates that a combination of impedance-based and HCS technologies provides a robust tool for in vitro assessment of CS constituents.

High Content Screening Analysis to Evaluate the Toxicological Effects of Harmful and Potentially Harmful Constituents HPHC

The objective of the study was to assess the biological impact of 15 cigarette smoke constituents using a combination of an impedance-based real time cell analyzer and a high-content screening (HCS)-based platform for toxicological assessment in vitro. This study provides information on effective doses, toxicity and modes of action of the tested compounds.

高内涵筛选分析法评价有害和潜在有害成分的毒性效应（HPHC）

Cigarette smoke (CS) is a major risk factor for cardiovascular and lung diseases. Because CS is a complex aerosol containing more than 7,000 chemicals1 it ...