Human Pluripotent Stem Cell Culture: Considerations for Maintenance, Expansion, and Therapeutics

Cell stem cell (Impact Factor: 22.27). 01/2014; 14(1):13-26. DOI: 10.1016/j.stem.2013.12.005
Source: PubMed


Human pluripotent stem cells (hPSCs) provide powerful resources for application in regenerative medicine and pharmaceutical development. In the past decade, various methods have been developed for large-scale hPSC culture that rely on combined use of multiple growth components, including media containing various growth factors, extracellular matrices, 3D environmental cues, and modes of multicellular association. In this Protocol Review, we dissect these growth components by comparing cell culture methods and identifying the benefits and pitfalls associated with each one. We further provide criteria, considerations, and suggestions to achieve optimal cell growth for hPSC expansion, differentiation, and use in future therapeutic applications.

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Available from: Kevin G Chen, Jun 12, 2014
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    • "However, in order to transform such proof-of-principle studies into viable therapeutic approaches for human TLE patients, it is critical to develop optimal human cell sources that can integrate into host circuitry and increase GABA-mediated inhibitory tone, thereby reducing seizure activity in the epileptic brain. Human pluripotent stem cell (hPSC) technologies, including induced PSCs (iPSCs), have the potential to provide an unlimited and ethically unimpeded source of therapeutic cells (Chen et al., 2014; Mallon et al., 2013; Yu et al., 2013) including human interneurons . Nevertheless, efficient translation of hPSC-derived interneurons could be hampered by their well-known, protracted maturation (Le Magueresse and Monyer, 2013; Nicholas et al., 2013). "
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    ABSTRACT: Seizure disorders debilitate more than 65,000,000 people worldwide, with temporal lobe epilepsy (TLE) being the most common form. Previous studies have shown that transplantation of GABA-releasing cells results in suppression of seizures in epileptic mice. Derivation of interneurons from human PSC has been reported, pointing to clinical translation of quality-controlled human cell sources that can enhance inhibitory drive and restore host circuitry. In this study, we demonstrate that human PSC-derived maturing GABAergic interneurons (mGIN) migrate extensively and integrate into dysfunctional circuitry of the epileptic mouse brain. Using optogenetic approaches, we find that grafted mGINs generate inhibitory postsynaptic responses in host hippocampal neurons. Importantly, even before acquiring full electrophysiological maturation, grafted neurons were capable of suppressing seizures and ameliorating behavioral abnormalities such as cognitive deficits, aggressiveness and hyperactivity. These results provide support for the potential of human PSC-derived mGIN for restorative cell therapy for epilepsy.
    Cell Stem Cell 11/2014; 15(5):559–573. DOI:10.1016/j.stem.2014.10.006 · 22.27 Impact Factor
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    • "These cells therefore represent powerful resources for applications in regenerative medicine and pharmaceutical development. However, several technical challenges must be addressed before hPSCs can be used routinely for clinical therapeutic applications and generation of tissues or organs [1]. First, in order to generate sufficient number of cells, culture systems that are cheap, easy-to handle and chemically defined are needed. "
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    ABSTRACT: Human pluripotent stem cells (hPSCs) can undergo unlimited self-renewal and have the capacity to differentiate into all somatic cell types, and are therefore an ideal source for the generation of cells and tissues for research and therapy. To realize this potential, defined cell culture systems that allow expansion of hPSCs and subsequent controlled differentiation, ideally in an implantable three-dimensional (3D) matrix, are required. Here we mimic spider silk - Nature's high performance material - for the design of chemically defined 2D and 3D matrices for cell culture. The silk matrices do not only allow xeno-free long-term expansion of hPSCs but also differentiation in both 2D and 3D. These results show that biomimetic spider silk matrices enable hPSC culture in a manner that can be applied for experimental and clinical purposes.
    Biomaterials 07/2014; 35(30). DOI:10.1016/j.biomaterials.2014.06.039 · 8.56 Impact Factor
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    • "Also, it is important to stress that established differentiation protocols will not necessarily transition seamlessly to a different culture system and can therefore represent a high barrier to progressing culture conditions even when long-term gains are significant. Different media used for the different systems have been thoroughly and recently reviewed elsewhere (Chen et al., 2014). Here, we will briefly describe selected defined media for feeder free culture . "
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    ABSTRACT: Self-renewing stem cell populations are increasingly considered as resources for cell therapy and tools for drug discovery. Human pluripotent stem (hPS) cells in particular offer a virtually unlimited reservoir of homogeneous cells and can be differentiated toward diverse lineages. Many diseases show impairment in self-renewal or differentiation, abnormal lineage choice or other aberrant cell behavior in response to chemical or physical cues. To investigate these responses, there is a growing interest in the development of specific assays using hPS cells, artificial microenvironments and high content analysis. Several hurdles need to be overcome that can be grouped into three areas: (i) availability of robust, homogeneous, and consistent cell populations as a starting point; (ii) appropriate understanding and use of chemical and physical microenvironments; (iii) development of assays that dissect the complexity of cell populations in tissues while mirroring specific aspects of their behavior. Here we review recent progress in the culture of hPS cells and we detail the importance of the environment surrounding the cells with a focus on synthetic material and suitable high content analysis approaches. The technologies described, if properly combined, have the potential to create a paradigm shift in the way diseases are modeled and drug discovery is performed.
    Frontiers in Pharmacology 07/2014; 5:150. DOI:10.3389/fphar.2014.00150 · 3.80 Impact Factor
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