A functionally characterized test set of human induced pluripotent stem cells

The Howard Hughes Medical Institute, Cambridge, Massachusetts, USA.
Nature Biotechnology (Impact Factor: 41.51). 02/2011; 29(3):279-86. DOI: 10.1038/nbt.1783
Source: PubMed


Human induced pluripotent stem cells (iPSCs) present exciting opportunities for studying development and for in vitro disease modeling. However, reported variability in the behavior of iPSCs has called their utility into question. We established a test set of 16 iPSC lines from seven individuals of varying age, sex and health status, and extensively characterized the lines with respect to pluripotency and the ability to terminally differentiate. Under standardized procedures in two independent laboratories, 13 of the iPSC lines gave rise to functional motor neurons with a range of efficiencies similar to that of human embryonic stem cells (ESCs). Although three iPSC lines were resistant to neural differentiation, early neuralization rescued their performance. Therefore, all 16 iPSC lines passed a stringent test of differentiation capacity despite variations in karyotype and in the expression of early pluripotency markers and transgenes. This iPSC and ESC test set is a robust resource for those interested in the basic biology of stem cells and their applications.

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Available from: John T Dimos, Oct 05, 2015
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    • "These precursors are in turn terminally differentiated into dopaminergic neurons under the effect of other signals (Fig. 1D). Similarly, spinal cord identity can be induced through the combinatorial addition of the 'caudal morphogens' RA, Wnts and FGFs (Amoroso et al., 2013; Boulting et al., 2011; Dimos et al., 3139 REVIEW Development (2015) 142, 3138-3150 doi:10.1242/dev.120568 DEVELOPMENT 2008; Li et al., 2005; Maury et al., 2015; Takazawa et al., 2012; Wichterle et al., 2002), again similar to the mechanisms operating in vivo (Liu et al., 2001; Nordström et al., 2002; Nordström et al., 2006). "
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    ABSTRACT: The human brain is arguably the most complex structure among living organisms. However, the specific mechanisms leading to this complexity remain incompletely understood, primarily because of the poor experimental accessibility of the human embryonic brain. Over recent years, technologies based on pluripotent stem cells (PSCs) have been developed to generate neural cells of various types. While the translational potential of PSC technologies for disease modeling and/or cell replacement therapies is usually put forward as a rationale for their utility, they are also opening novel windows for direct observation and experimentation of the basic mechanisms of human brain development. PSC-based studies have revealed that a number of cardinal features of neural ontogenesis are remarkably conserved in human models, which can be studied in a reductionist fashion. They have also revealed species-specific features, which constitute attractive lines of investigation to elucidate the mechanisms underlying the development of the human brain, and its link with evolution.
    Development 09/2015; 142(18):3138-3150. DOI:10.1242/dev.120568 · 6.46 Impact Factor
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    • "The issue regarding the use of differentiation protocols remains unclear. Similar differentiation efficiencies have been obtained by independent laboratories using the same standardised procedures (Boulting et al., 2011), supporting the reproducibility of iPSC-based studies. However , many iPSC-based studies use alternative differentiation protocols, which make it difficult to interpret findings from different studies. "
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    ABSTRACT: Over recent years tremendous progress has been made towards understanding the molecular and cellular mechanism by which estrogens exert enhancing effects on cognition, and how they act as a neuroprotective or neurotrophic agent in disease. Currently, much of this work has been carried out in animal models with only a limited number of studies using native human tissue or cells. Recent advances in stem cell technology now make it possible to reprogram somatic cells from humans into induced pluripotent stem cells (iPSCs), which can subsequently be differentiated in neurons of specific lineages. Importantly, the reprogramming of cells allows for the generation of iPSCs that retains the genetic "makeup" of the donor. Therefore, it is possible to generate iPSC-derived neurons from patients diagnosed with specific diseases, that harbor the complex genetic background associated with the disorder. Here, we review the iPSC technology and how its currently being used to model neural development and neurological diseases. Furthermore, we explore whether this cellular system could be used to understand the role of estrogens in human neurons, and present preliminary data in support of this. We further suggest that the use of iPSC technology offers a novel system in which to not only further understand estrogens' effects in human cells, but in which to investigate the mechanism by which estrogens are beneficial in disease. Developing a greater understanding of these mechanisms in native human cells will also aid in the development of safer and more effective estrogen-based therapeutics. Copyright © 2015. Published by Elsevier Inc.
    Hormones and Behavior 07/2015; 26. DOI:10.1016/j.yhbeh.2015.06.014 · 4.63 Impact Factor
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    • "Third, the proportion of diseaserelevant motor neuron subtypes remains most often undetermined in human iPSc-derived cultures (Bilican et al., 2012; Boulting et al., 2011; Egawa et al., 2012; Hester et al., 2011; Karumbayaram et al., 2009; Mitne-Neto et al., 2011; Qu et al., 2014; Serio et al., 2013) but see Amoroso et al. (2013). Fourth, the presence of unwanted cell types such as neural precursors, various types of neurons (Amoroso et al., 2013; Boulting et al., 2011; Chen et al., 2014; Dimos et al., 2008; Ebert et al., 2009; Karumbayaram et al., 2009; Naujock et al., 2014; Qu et al., 2014; Sareen et al., 2012) or astrocytes (Haidet-Phillips et al., 2011; Re et al., 2014) may induce confounding effects on motor neuron survival. Likewise, pharmacological compounds may act indirectly through these cell types rendering difficult the interpretation of their effects. "
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    ABSTRACT: Amyotrophic lateral sclerosis (ALS) is a severe and incurable neurodegenerative disease. Human motor neurons generated from induced pluripotent stem cells (iPSc) offer new perspectives for disease modeling and drug testing in ALS. In standard iPSc-derived cultures however, the two major phenotypic alterations of ALS - degeneration of motor neuron cell bodies and axons - are often obscured by cell body clustering, extensive axon criss-crossing and presence of unwanted cell types. Here, we succeeded to isolate 100% pure and standardized human motor neurons by a novel FACS double selection based on a p75(NTR) surface epitope and an HB9:RFP lentivirus reporter. The p75(NTR)/HB9::RFP motor neurons survive and grow well without forming clusters or entangled axons, are electrically excitable, contain ALS-relevant motor neuron subtypes and form functional connections with co-cultured myotubes. Importantly, they undergo rapid and massive cell death and axon degeneration in response to mutant SOD1 astrocytes. These data demonstrate the potential of FACS-isolated human iPSc-derived motor neurons for improved disease modeling and drug testing in ALS and related motor neuron diseases. Copyright © 2015. Published by Elsevier Inc.
    Neurobiology of Disease 06/2015; 82. DOI:10.1016/j.nbd.2015.06.011 · 5.08 Impact Factor
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