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.
Highlights
HUMAN PLURIPOTENT STEM CELLS ARE DERIVED FROM EMBRYOS OR THROUGH REPROGRAMMING Stem cells are defined as cells capable of self-renewal, the capacity to generate identical copies of themselves, and differentiation, the ability to provide cells performing a specific biological function (Smith, 2006).The capacity to differentiate into all lineages sufficient to form an entire organism, and not necessarily extra embryonic tissues, is defined as pluripotency
Several types of cells have been characterized which broadly fit many aspects of the definition of pluripotency. HUMAN PLURIPOTENT STEM (hPS) cells are derived from transient populations of cells isolated from the embryo such as human embryonic stem cells or are artificially reprogrammed from somatic cells such as human induced pluripotent stem cells
Phenotyping hPSCs on artificial microenvironments system, cell culture is artificial by definition and it is not easy to pinpoint what the “natural” conditions are in vivo and should be in vitro
Summary
HUMAN PLURIPOTENT STEM (hPS) CELLS ARE DERIVED FROM EMBRYOS OR THROUGH REPROGRAMMING Stem cells are defined as cells capable of self-renewal, the capacity to generate identical copies of themselves, and differentiation, the ability to provide cells performing a specific biological function (Smith, 2006).The capacity to differentiate into all lineages sufficient to form an entire organism, and not necessarily extra embryonic tissues, is defined as pluripotency. DEFINING THE CULTURE: FEEDERS AND MEDIA Irrespective of the biological differences, expansion of homogeneous starting populations in self-renewing conditions is key to realizing the promise of hPS cells for screening and modeling strategies.
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