Recommendations, guidelines, and best practice for the use of human induced pluripotent stem cells for neuropharmacological studies of neuropsychiatric disorders

Polit, Lucia Dutan; Eidhof, Ilse; McNeill, Rhiannon V.; Warre-Cornish, Katherine; Ohki, Cristine Marie Yde; Walter, Natalie Monet; Sala, Carlo; Verpelli, Chiara; Radtke, Franziska; Galderisi, Silvana; Mucci, Armida; Collo, Ginetta; Edenhofer, Frank; Castrén, Maija; Réthelyi, János; Ejlersen, Morten; Hohmann, Sonja; Ilieva, Mirolyuba; Lukjanska, Renate; Matuleviciute, Rugile; Michel, Tanja Maria; Vrij, Femke M.S. de; Kushner, Steven A.; Lendemeijer, Bas; Kittel‐Schneider, Sarah; Ziegler, Georg C.; Gruber‐Schoffnegger, Doris; Pasterkamp, R. Jeroen; Kasri, Amal; Potier, Marie‐Claude; Knoblich, Jürgen A.; Brüstle, Oliver; Peitz, Michael; Pich, Emilio Merlo; Harwood, Adrian J.; Abranches, Elsa; Falk, Anna; Vernon, Anthony C.; Grünblatt, Edna; Srivastava, Deepak P.

doi:10.1016/j.nsa.2023.101125

Cited by 9 publications

(4 citation statements)

References 182 publications

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“…Fourth, the noticeable presence of variation between different donor lines may mask detectable differences at an averaged donor level. Our sample size of N=4 donor lines, although in line with recommendations for the minimum sample size in such experiments (Dutan Polit et al, 2023), coupled with the variability in responses generated by each individual donor may well constrain our ability to detect group-level differences. This is, however, consistent with the known heterogeneity of outcomes in response to maternal immune activation in humans and in rodent models, and that in most cases there is no negative outcome (Meyer, 2019;Mueller et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

“…Fourth, the noticeable presence of variation between different donor lines may mask detectable differences at an averaged donor level. Our sample size of N=4 donor lines, although in line with recommendations for the minimum sample size in such experiments (Dutan Polit et al, 2023), coupled with the variability in responses generated by each individual donor may well constrain our ability to detect group-level differences. Additionally, the diversity among our donor samples may be influenced by sex, as illustrated in Figure 3A PCA, where the sole female line (069_CTF) separates from the three male lines along PC1.…”

Section: Discussionmentioning

confidence: 99%

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Transcriptional and Cellular Response of hiPSC-derived Microglia-Neural Progenitor Co-Cultures Exposed to IL-6

Couch,

Brown,

Raimundo

et al. 2023

Preprint

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Elevated interleukin (IL-)6 levels during prenatal development have been linked to increased risk for neurodevelopmental disorders (NDD) in the offspring, but the mechanism remains unclear. Human-induced pluripotent stem cell (hiPSC) models offer a valuable tool to study the effects of IL-6 on features relevant for human neurodevelopmentin vitro. We previously reported that hiPSC-derived microglia-like cells (MGLs) respond to IL-6, but neural progenitor cells (NPCs) in monoculture do not. We therefore investigated whether co-culturing hiPSC-derived MGLs with NPCs would trigger a cellular response to IL-6 stimulation via secreted factors from the MGLs. Using N=4 donor lines without psychiatric diagnosis, we first confirmed that NPCs can respond to IL-6 through trans-signalling when recombinant IL-6Ra is present, and that this response is dose-dependent. In response to IL-6, MGLs secreted soluble IL-6R, but at lower levels than found in vivo and below that needed to activate trans-signalling in NPCs. Among other cytokines, MGLs also increased Tumour necrosis factor (TNF)α secretion into the co-culture environment. Despite this, whilst transcriptomic analysis confirmed that MGLs undergo substantial transcriptomic changes after IL-6 exposure, NPCs in co-culture with MGLs exhibited a minimal transcriptional response. Furthermore, there were no significant cell fate-acquisition changes when differentiated into post-mitotic cultures, nor alterations in synaptic densities in mature neurons. These findings highlight the need to investigate if trans-IL-6 signalling to NPCs is a relevant disease mechanism linking prenatal IL-6 exposure to increased risk for psychiatric disorders. Moreover, our findings underscore the importance of establishing more complexin vitrohuman models with diverse cell types, which may show cell-specific responses to microglia-released cytokines to fully understand how IL-6 exposure may influence human neurodevelopment.

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Section: Discussionmentioning

confidence: 99%

“…Fourth, the noticeable presence of variation between different donor lines may mask detectable differences at an averaged donor level. Our sample size of N=4 donor lines, although in line with recommendations for the minimum sample size in such experiments (Dutan Polit et al, 2023), coupled with the variability in responses generated by each individual donor may well constrain our ability to detect group-level differences. Additionally, the diversity among our donor samples may be influenced by sex, as illustrated in Figure 3A PCA, where the sole female line (069_CTF) separates from the three male lines along PC1.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional and Cellular Response of hiPSC-derived Microglia-Neural Progenitor Co-Cultures Exposed to IL-6

Couch,

Brown,

Raimundo

et al. 2023

Preprint

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“…Despite all the relevant advantages, the experimental reproducibility remains one of the main challenges in hiPSC-based research, and must be taken into account: approaches to limit variability in protocols and data need to be addressed. It is mainly due to: genetic heterogeneity from donors, reprogramming methods of the isolation source (skin or blood cells), changes induced during passages in culture and differentiation protocols (Ardhanareeswaran et al, 2017;Dutan Polit et al, 2023;Hong et al, 2023).…”

Section: Induced Pluripotent Stem Cellsmentioning

confidence: 99%

Advanced materials and biofabrication technologies to design in vitro functional central nervous system models

Traldi,

Chiappini,

Menduti

et al. 2023

Front. Med. Eng.

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Nowadays, the pathophysiology of several central nervous system (CNS) disorders is still poorly understood, making difficult the identification of efficient treatments. CNS damages, due to neurodegenerative conditions or injuries, often result in permanent neuronal dysfunctions and serious impairments of motor, sensory and cognitive capacities. Despite the many attempts of pharmaceutical research to promote neural regeneration, poor progresses have been made in effectively restoring nervous functionality. Indeed, most of the experimental drugs show limited efficacy in the clinical trials, also due to existing preclinical models’ inability in fully replicating the complexity of CNS pathophysiology. Therefore, tissue-engineered three-dimensional (3D) models are being extensively explored to develop novel representative in vitro platforms, which more carefully replicate the architecture of neural microenvironment, including both cellular and extracellular components. In this respect, 3D in vitro models are expected to be promising and comprehensive tools for investigating CNS diseases and testing new drug compounds, as they overcome some of the common limitations of traditional two-dimensional (2D) cultures. This review discusses the main challenges to be addressed in CNS modeling, analyzing the key elements involved in neural tissue engineering. Specifically, an overview of the mostly used neural cell sources and biomaterials is provided, focusing on the critical aspects to consider in selecting the appropriate components according to the application. Different methods adopted to modulate the structural and functional properties of the engineered microenvironment are also presented, aimed at fostering in vitro tissue maturation. Lastly, the latest advances in biofabrication technologies are outlined, reviewing the most recent 3D bioprinted in vitro systems and microfluidic-based 3D platforms, starting from the modeling of distinctive CNS pathophysiological mechanisms to the designing of refined and functional in vivo-like neural microtissues.

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“…High culture costs associated with undefined iPSC matrices and media further limit their scalability 2,3 . To address all these challenges, recent efforts have been made by large networks, like the International Society for Stem Cell Research (ISSCR) (https://www.isscr.org/standards-document) and the European College of Neuropsychopharmacology (ECNP) Network to provide researchers with best practices and recommendations for the use of human stem cells in research 4 . Moreover, researchers have explored xeno-free, fully defined iPSC culture matrices, such as laminin 521 (LN-521) and vitronectin, and defined iPSC culture media, such as essential 8 (E8) to facilitate the robust generation and maintenance of iPSCs 2,3,5 .…”

Section: Introductionmentioning

confidence: 99%

Defined human PSC culture conditions robustly maintain human PSC pluripotency through Ca²⁺signaling

Eidhof

Kele

Shahsavani

et al. 2023

Preprint

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Human pluripotent stem cells (hPSCs) have significant potential for disease modeling and cell therapies. However, their clinical applicability is limited due to the need for undefined conditions for PSC cultivation, which increase the risk of immunogenicity, result in batch-to-batch variability and finite scalability. These limitations may be circumvented by xeno-free, defined culture conditions. However, biological processes that preserve robust, homogenous PSCs in defined conditions remain to be characterized. Here, we compared gene expression data from over 100 hPSC cell lines cultivated in undefined and defined culture conditions. Defined culture conditions significantly reduced inter-PSC line variability, highlighting the importance of standardization to minimize PSC biases. This variability is concurrent with decreased germ layer differentiation and increased expression of Ca2+-binding proteins. The significance of tightly controlled Ca2+ signaling in hPSC pluripotency in defined culture conditions was also confirmed. A deeper understanding of these processes may aid in standardizing defined hPSC culture conditions.

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Recommendations, guidelines, and best practice for the use of human induced pluripotent stem cells for neuropharmacological studies of neuropsychiatric disorders

Cited by 9 publications

References 182 publications

Transcriptional and Cellular Response of hiPSC-derived Microglia-Neural Progenitor Co-Cultures Exposed to IL-6

Transcriptional and Cellular Response of hiPSC-derived Microglia-Neural Progenitor Co-Cultures Exposed to IL-6

Advanced materials and biofabrication technologies to design in vitro functional central nervous system models

Defined human PSC culture conditions robustly maintain human PSC pluripotency through Ca²⁺signaling

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Recommendations, guidelines, and best practice for the use of human induced pluripotent stem cells for neuropharmacological studies of neuropsychiatric disorders

Cited by 9 publications

References 182 publications

Transcriptional and Cellular Response of hiPSC-derived Microglia-Neural Progenitor Co-Cultures Exposed to IL-6

Transcriptional and Cellular Response of hiPSC-derived Microglia-Neural Progenitor Co-Cultures Exposed to IL-6

Advanced materials and biofabrication technologies to design in vitro functional central nervous system models

Defined human PSC culture conditions robustly maintain human PSC pluripotency through Ca2+signaling

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Product

Resources

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Defined human PSC culture conditions robustly maintain human PSC pluripotency through Ca²⁺signaling