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Complex Tissue and Disease Modeling using hiPSCs

Robert Passier, Valeria Orlova, Christine Mummery
2016 Cell Stem Cell  
CONFLICTS OF INTEREST Christine Mummery and Robert Passier are co-founders of Pluriomics bv. Christine Mummery is on the advisory board of Galapagos bv.  ...  There is particular potential here for hiPSCs since these are now amenable to stable integration of reporter constructs (Den Hartogh and Passier, 2016) .  ... 
doi:10.1016/j.stem.2016.02.011 pmid:26942851 fatcat:sxlq26isqjcazm7vetcaxiqyta

Native cardiac environment and its impact on engineering cardiac tissue

Verena Schwach, Robert Passier
2019 Biomaterials Science  
In this review, we describe the progressive build-up of the cardiac extracellular matrix (ECM) during embryonic development, the ECM of the adult human heart and the application of natural and synthetic biomaterials for cardiac tissue engineering using hPSC-CMs.
doi:10.1039/c8bm01348a pmid:31338495 fatcat:466dfnzdfrg6neu634jzbd636q

Getting to the Heart of the Matter: Direct Reprogramming to Cardiomyocytes

Robert Passier, Christine Mummery
2010 Cell Stem Cell  
Although many hurdles remain to be overcome in order to establish effective cellbased therapies (Passier et al., 2008) , patient-derived iCMs may be used in the future for treatment of heart disease and  ... 
doi:10.1016/j.stem.2010.07.004 pmid:20682439 fatcat:bqimzliwczgujpiqirjx57z6na

Human Pluripotent Stem Cell-Derived Cardiomyocytes for Assessment of Anticancer Drug-Induced Cardiotoxicity

Verena Schwach, Rolf H. Slaats, Robert Passier
2020 Frontiers in Cardiovascular Medicine  
Schöffski P, Delord JPP, Brain E, Robert J, Dumez H, Gasmi J, et al.  ... 
doi:10.3389/fcvm.2020.00050 pmid:32322588 pmcid:PMC7156610 fatcat:k7eocdkfujf37cfsn3oufqghsa

Personalised organs-on-chips: functional testing for precision medicine

Albert van den Berg, Christine L. Mummery, Robert Passier, Andries D. van der Meer
2019 Lab on a Chip  
Organs-on-chips can be 'personalised' so they can be used as functional tests to inform clinical decision-making for specific patients.
doi:10.1039/c8lc00827b pmid:30506070 pmcid:PMC6336148 fatcat:gxohu7gqjndsvhhcfzoqtkra74

Automated image analysis system for studying cardiotoxicity in human pluripotent stem cell-Derived cardiomyocytes

Lu Cao, Andries D. van der Meer, Fons J. Verbeek, Robert Passier
2020 BMC Bioinformatics  
Cardiotoxicity, characterized by severe cardiac dysfunction, is a major problem in patients treated with different classes of anticancer drugs. Development of predictable human-based models and assays for drug screening are crucial for preventing potential drug-induced adverse effects. Current animal in vivo models and cell lines are not always adequate to represent human biology. Alternatively, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) show great potential for
more » ... se modelling and drug-induced toxicity screenings. Fully automated high-throughput screening of drug toxicity on hiPSC-CMs by fluorescence image analysis is, however, very challenging, due to clustered cell growth patterns and strong intracellular and intercellular variation in the expression of fluorescent markers. In this paper, we report on the development of a fully automated image analysis system for quantification of cardiotoxic phenotypes from hiPSC-CMs that are treated with various concentrations of anticancer drugs doxorubicin or crizotinib. This high-throughput system relies on single-cell segmentation by nuclear signal extraction, fuzzy C-mean clustering of cardiac α-actinin signal, and finally nuclear signal propagation. When compared to manual segmentation, it generates precision and recall scores of 0.81 and 0.93, respectively. Our results show that our fully automated image analysis system can reliably segment cardiomyocytes even with heterogeneous α-actinin signals.
doi:10.1186/s12859-020-3466-1 pmid:32408861 fatcat:2sfu4de6lndonlci63sdv4jtfa

CHAMP, A Novel Cardiac-Specific Helicase Regulated by MEF2C

Zhi-Ping Liu, Osamu Nakagawa, Masayo Nakagawa, Hiromi Yanagisawa, Robert Passier, James A. Richardson, Deepak Srivastava, Eric N. Olson
2001 Developmental Biology  
MEF2C is a MADS-box transcription factor required for cardiac myogenesis and morphogenesis. In MEF2C mutant mouse embryos, heart development arrests at the looping stage (embryonic day 9.0), the future right ventricular chamber fails to form, and cardiomyocyte differentiation is disrupted. To identify genes regulated by MEF2C in the developing heart, we performed differential array analysis coupled with subtractive cloning using RNA from heart tubes of wild-type and MEF2C-null embryos. Here, we
more » ... describe a novel MEF2C-dependent gene that encodes a cardiac-restricted protein, called CHAMP (cardiac helicase activated by MEF2 protein), that contains seven conserved motifs characteristic of helicases involved in RNA processing, DNA replication, and transcription. During mouse embryogenesis, CHAMP expression commences in the linear heart tube at embryonic day 8.0, shortly after initiation of MEF2C expression in the cardiogenic region. Thereafter, CHAMP is expressed specifically in embryonic and postnatal cardiomyocytes. At the trabeculation stage of heart development, CHAMP expression is highest in the trabecular region in which cardiomyocytes have exited the cell cycle and is lowest in the proliferative compact zone. These findings suggest that CHAMP acts downstream of MEF2C in a cardiac-specific regulatory pathway for RNA processing and/or transcriptional control.
doi:10.1006/dbio.2001.0277 pmid:11397016 fatcat:jk5sjjes5ffohcbtk2wslyamzi

Cardiac differentiation of pluripotent stem cells and implications for modeling the heart in health and disease

Harsha D. Devalla, Robert Passier
2018 Science Translational Medicine  
doi:10.1126/scitranslmed.aah5457 pmid:29618562 fatcat:5yxliskaqbgk5mr5uoi4gsr5zm

Membranes for Modelling Cardiac Tissue Stiffness In Vitro Based on Poly(trimethylene carbonate) and Poly(ethylene glycol) Polymers

Iris Allijn, Marcelo Ribeiro, André Poot, Robert Passier, Dimitrios Stamatialis
2020 Membranes  
Despite the increased expenditure of the pharmaceutical industry on research and development, the number of drugs for cardiovascular diseases that reaches the market is decreasing. A major issue is the limited ability of the current in vitro and experimental animal models to accurately mimic human heart disease, which hampers testing of the efficacy of potential cardiac drugs. Moreover, many non-heart-related drugs have severe adverse cardiac effects, which is a major cause of drugs' retraction
more » ... after approval. A main hurdle of current in vitro models is their inability to mimic the stiffness of in vivo cardiac tissue. For instance, poly(styrene) petri dishes, which are often used in these models, have a Young's modulus in the order of GPa, while the stiffness of healthy human heart tissue is <50 kPa. In pathological conditions, such as scarring and fibrosis, the stiffness of heart tissue is in the >100 kPa range. In this study, we focus on developing new membranes, with a set of properties for mimicry of cardiac tissue stiffness in vitro, based on methacrylate-functionalized macromers and triblock-copolymers of poly(trimethylene carbonate) and poly(ethylene glycol). The new membranes have Young's moduli in the hydrated state ranging from 18 kPa (healthy tissue) to 2.5 MPa (pathological tissue), and are suitable for cell contraction studies using human pluripotent stem-cell-derived cardiomyocytes. The membranes with higher hydrophilicity have low drug adsorption and low Young's moduli and could be suitable for drug screening applications.
doi:10.3390/membranes10100274 pmid:33022962 pmcid:PMC7650615 fatcat:ga4mnznvonbernbf6ba2xpljtm

Human Pluripotent Stem Cell Differentiation into Functional Epicardial Progenitor Cells

Juan Antonio Guadix, Valeria V. Orlova, Elisa Giacomelli, Milena Bellin, Marcelo C. Ribeiro, Christine L. Mummery, José M. Pérez-Pomares, Robert Passier
2017 Stem Cell Reports  
Multiple cell-based strategies to regenerate damaged post-mitotic tissues have been described (reviewed by Passier et al., 2008) .  ... 
doi:10.1016/j.stemcr.2017.10.023 pmid:29173898 pmcid:PMC5785703 fatcat:vh7qzp5djffb3cgzqygj6xplpi

Prediction of drug-induced cardiotoxicity using human embryonic stem cell-derived cardiomyocytes

Stefan R. Braam, Leon Tertoolen, Anja van de Stolpe, Thomas Meyer, Robert Passier, Christine L. Mummery
2010 Stem Cell Research  
., 2003; Passier et al., 2008; Satin et al., 2004) .  ... 
doi:10.1016/j.scr.2009.11.004 pmid:20034863 fatcat:45d7n7dqmfb5bdhtwmabbzilee

Fluidic Circuit Board with Modular Sensor and Valves Enables Stand-Alone, Tubeless Microfluidic Flow Control in Organs-on-Chips [article]

Aisen Vivas, Albert van den Berg, Robert Passier, Mathieu Odijk, Andries D. van der Meer
2021 bioRxiv   pre-print
Organs-on-chips are a unique class of microfluidic in vitro cell culture models, in which the in vivo tissue microenvironment is mimicked. Unfortunately, its widespread use is hampered by their operation complexity and incompatibility with end-user research settings. To address these issues, many commercial and non-commercial platforms have been developed for semi-automated culture of organs-on-chips. However, these organ-on-chip culture platforms each represent a closed ecosystem, with very
more » ... tle opportunity to interchange and integrate components from different platforms or to develop new ones. The Translational Organ-on-Chip Platform (TOP) is a multi-institutional effort to develop an open platform for automated organ-on-chip culture and integration of components from various developers. Central to TOP is the fluidic circuit board (FCB), a microfluidic plate with the form factor of a typical well plate. The FCB enables microfluidic control of multiple components like sensors or organ-on-chip devices through an interface based on openly available standards. Here, we report an FCB to integrate commercial and in-house developed components forming a stand-alone flow control system for organs-on-chips. The control system is able to achieve constant and pulsatile flow recirculation through a connected organ-on-chip device. We demonstrate that this system is able to automatically perfuse a heart-on-chip device containing co-cultures of cardiac tissues derived from human pluripotent stem cell-derived cardiomyocytes and monolayers of endothelial cells for five days. Altogether, we conclude that open technology platforms allow the integration of components from different sources to form functional and fit-for-purpose organ-on-chip systems. We anticipate that open platforms will play a central role in catalysing and maturing further technological development of organ-on-chip culture systems.
doi:10.1101/2021.11.24.469685 fatcat:bcy5uyw7gzbs7ima6xk7ykn7pq

A Quest for Human and Mouse Embryonic Stem Cell-specific Proteins

Dennis Van Hoof, Robert Passier, Dorien Ward-Van Oostwaard, Martijn W. H. Pinkse, Albert J. R. Heck, Christine L. Mummery, Jeroen Krijgsveld
2006 Molecular & Cellular Proteomics  
Embryonic stem cells (ESCs) are of immense interest as they can proliferate indefinitely in vitro and give rise to any adult cell type, serving as a potentially unlimited source for tissue replacement in regenerative medicine. Extensive analyses of numerous human and mouse ESC lines have shown generic similarities and differences at both the transcriptional and functional level. However, comprehensive proteome analyses are missing or are restricted to mouse ESCs. Here we have used an extensive
more » ... roteomic approach to search for ESC-specific proteins by analyzing the differential protein expression profiles of human and mouse ESCs and their differentiated derivatives. The data sets comprise 1,775 non-redundant proteins identified in human ESCs, 1,532 in differentiated human ESCs, 1,871 in mouse ESCs, and 1,552 in differentiated mouse ESCs with a false positive rate of <0.2%. Comparison of the data sets distinguished 191 proteins exclusively identified in both human and mouse ESCs but not in their differentiated derivatives. Besides well known ESC benchmarks, this subset included many uncharacterized proteins, some of which may be novel ESC-specific markers. To complement the mass spectrometric approach, differential expression of a selection of these proteins was confirmed by Western blotting, immunofluorescence confocal microscopy, and fluorescence-activated cell sorting. Additionally two other independently isolated and cultured human ESC lines as well as their differentiated derivatives were monitored for differential expression of selected proteins. Some of these proteins were identified exclusively in ESCs of all three human lines and may thus serve as generic ESC markers. Our wide scale proteomic approach enabled us to screen thousands of proteins rapidly and select putative ESC-associated proteins for further analysis. Validation by three independent conventional protein analysis techniques shows that our methodology is robust, provides an excellent tool to char-acterize ESCs at the protein level, and may disclose novel ESC-specific benchmarks.
doi:10.1074/mcp.m500405-mcp200 pmid:16600995 fatcat:uuwqkrieznbknkindjhle5vexu

Modulation of Cardiac Growth and Development by HOP, an Unusual Homeodomain Protein

Chong Hyun Shin, Zhi-Ping Liu, Robert Passier, Chun-Li Zhang, Da-Zhi Wang, Thomas M. Harris, Hiroyuki Yamagishi, James A. Richardson, Geoffrey Childs, Eric N. Olson
2002 Cell  
In situ reproducibility in dye swap arrays, and a minimum median signal hybridization of whole embryos or paraffin sections was performed intensity of twice local background. as described (Passier et  ... 
doi:10.1016/s0092-8674(02)00933-9 pmid:12297046 fatcat:2qrlrmvfw5bajf4uyz6sjdxq5u

A comprehensive gene expression analysis at sequential stages of in vitro cardiac differentiation from isolated MESP1-expressing-mesoderm progenitors

Sabine C. den Hartogh, Katherine Wolstencroft, Christine L. Mummery, Robert Passier
2016 Scientific Reports  
In vitro cardiac differentiation of human pluripotent stem cells (hPSCs) closely recapitulates in vivo embryonic heart development, and therefore, provides an excellent model to study human cardiac development. We recently generated the dual cardiac fluorescent reporter MESP1 mCherry/w NKX2-5 eGFP/w line in human embryonic stem cells (hESCs), allowing the visualization of pre-cardiac MESP1+ mesoderm and their further commitment towards the cardiac lineage, marked by activation of the cardiac
more » ... nscription factor NKX2-5. Here, we performed a comprehensive whole genome based transcriptome analysis of MESP1-mCherry derived cardiac-committed cells. In addition to previously described cardiac-inducing signalling pathways, we identified novel transcriptional and signalling networks indicated by transient activation and interactive network analysis. Furthermore, we found a highly dynamic regulation of extracellular matrix components, suggesting the importance to create a versatile niche, adjusting to various stages of cardiac differentiation. Finally, we identified cell surface markers for cardiac progenitors, such as the Leucine-rich repeat-containing G-protein coupled receptor 4 (LGR4), belonging to the same subfamily of LGR5, and LGR6, established tissue/cancer stem cells markers. We provide a comprehensive gene expression analysis of cardiac derivatives from pre-cardiac MESP1-progenitors that will contribute to a better understanding of the key regulators, pathways and markers involved in human cardiac differentiation and development. HPSCs provide an excellent platform to model human heart development and cardiac differentiation in vitro. We and others have previously shown that there are close similarities between the temporal and sequential transcriptional activation of genes during human cardiac differentiation in vitro and in cardiac development in vivo 1,2 . Identification of key molecular events during cardiac differentiation from human pluripotent stem cells is instrumental for a better understanding of human cardiac development, and advancing the fields of regenerative medicine, disease modelling, and drug discovery. Although cardiac differentiation protocols have been improved significantly over the last years, in-depth knowledge of molecular mechanisms involved in cardiac lineage commitment is sparse. Specification of hPSCs towards the cardiac lineage is regulated by the precise temporal expression of transcriptional networks. Detailed information on these transcriptional networks and signalling molecules is essential for understanding the mechanisms underlying progression and expansion of progenitors and their differentiation to their specific subtype derivatives. When hPSCs differentiate towards the cardiac lineage, they first progress via an intermediate mesodermal stage, expressing transcription factor MESP1, before they further differentiate into cardiac derivatives, indicated by the increased expression of cardiac marker NKX2-5 3,4 . In order to understand how molecular mechanisms play a role in these lineage decisions, we previously generated the dual cardiac human embryonic stem cell (hESC) reporter line MESP1 mCherry/w NKX2-5 eGFP/w4 . We characterised MESP1 progenitors by gene expression and surface marker analysis, and showed that MESP1 derivatives were predominantly enriched for NKX2-5 positive cardiomyocytes. Here, we have defined the temporal gene expression changes that occur during the differentiation of isolated MESP1-progenitors towards their cardiac derivatives. By classifying enriched genes according to similar expression patterns, functional pathway analysis, GO terms, sequence-specific DNA binding properties, and protein-protein interactions, we provide more insight on molecular signals and regulators involved in the sequential stages of mesoderm to cardiac differentiation. We confirm the enrichment of previously identified transcriptional regulators and cardiac functional genes, indicated by the predominant activation of components of the Wnt and TGFβ pathways. Interestingly, we identified novel cardiac networks, signalling pathways and markers, which may have an important role in the early cardiac differentiation and may be used as blueprint to evaluate or study human cardiac differentiation in vitro.
doi:10.1038/srep19386 pmid:26783251 pmcid:PMC4726039 fatcat:3pezeciltrdqhdyifkvuyhsj6m
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