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      The likely role of proteolytic enzymes in unwanted differentiation of stem cells in culture

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          Abstract

          Tissue engineering aims at developing the necessary technological strategies for replacement or regeneration tissues. However, the number of cells required is much greater than the number obtained from a cell source. Expanding the cells' number in cell culture for a long period is required until the necessary amount of cells is obtained. While in culture, cells often go unwanted differentiation. Little attention has been given to the use of proteolytic enzymes in cell passage. Review the importance of extracellular matrix and surface proteins for cell behavior and the possible mechanisms of cellular changes that may occur due to the use of proteolytic enzymes is an essential issue. Possible alternative to replace these enzymes in cell passage has also been developed.

          Abstract

          Tissue engineering aims at developing strategies for tissue replacement. However, the number of cells required is much greater than the number obtained from a cell source. Expanding the cells' number in cell culture for a long period is required until the necessary amount of cells is obtained but, cells often go unwanted differentiation. Proteolytic enzymes are commonly used in culture and it might lead to such cellular changes. This article aims at reviewing the possible mechanisms of cellular changes that may occur due to the use of proteolytic enzymes and possible alternatives to replace the use of those enzymes.

          Most cited references34

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          Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer.

          The microenvironment influences gene expression so that the behavior of a cell is largely determined by its interactions with the extracellular matrix, neighboring cells, and soluble local and systemic cues. We describe the essential roles of context and organ structure in directing mammary gland development and differentiated function and in determining the response to oncogenic insults, including mutations. We expand on the concept of "dynamic reciprocity" to present an integrated view of development, cancer, and aging and posit that genes are like the keys on a piano: Although they are essential, it is the context that makes the music.
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            Global transcription in pluripotent embryonic stem cells.

            The molecular mechanisms underlying pluripotency and lineage specification from embryonic stem cells (ESCs) are largely unclear. Differentiation pathways may be determined by the targeted activation of lineage-specific genes or by selective silencing of genome regions. Here we show that the ESC genome is transcriptionally globally hyperactive and undergoes large-scale silencing as cells differentiate. Normally silent repeat regions are active in ESCs, and tissue-specific genes are sporadically expressed at low levels. Whole-genome tiling arrays demonstrate widespread transcription in coding and noncoding regions in ESCs, whereas the transcriptional landscape becomes more discrete as differentiation proceeds. The transcriptional hyperactivity in ESCs is accompanied by disproportionate expression of chromatin-remodeling genes and the general transcription machinery. We propose that global transcription is a hallmark of pluripotent ESCs, contributing to their plasticity, and that lineage specification is driven by reduction of the transcribed portion of the genome.
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              Engineering hydrogels as extracellular matrix mimics.

              Extracellular matrix (ECM) is a complex cellular environment consisting of proteins, proteoglycans, and other soluble molecules. ECM provides structural support to mammalian cells and a regulatory milieu with a variety of important cell functions, including assembling cells into various tissues and organs, regulating growth and cell-cell communication. Developing a tailored in vitro cell culture environment that mimics the intricate and organized nanoscale meshwork of native ECM is desirable. Recent studies have shown the potential of hydrogels to mimic native ECM. Such an engineered native-like ECM is more likely to provide cells with rational cues for diagnostic and therapeutic studies. The research for novel biomaterials has led to an extension of the scope and techniques used to fabricate biomimetic hydrogel scaffolds for tissue engineering and regenerative medicine applications. In this article, we detail the progress of the current state-of-the-art engineering methods to create cell-encapsulating hydrogel tissue constructs as well as their applications in in vitro models in biomedicine.
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                Author and article information

                Journal
                Future Sci OA
                Future Sci OA
                FSO
                Future Science OA
                Future Science Ltd (London, UK )
                2056-5623
                November 2015
                01 November 2015
                : 1
                : 3
                : FSO28
                Affiliations
                [1 ]Graduate Translational Surgery Program, Paulista School of Medicine, Federal University of Sao Paulo (EPM/UNIFESP), SP, Brazil
                [2 ]CTCMol, Center of Cellular & Molecular Therapy, São Paulo, SP, Brazil
                [3 ]Department of Morphology & Basic Pathology, Jundiaí Faculty of Medicine, São Paulo, SP, Brazil
                [4 ]Medicine Department, Endocrinology Division, EPM/UNIFESP, Sao Paulo, SP, Brazil
                [5 ]Surgery Department, Translational Surgery Division, EPM/UNIFESP, Sao Paulo, SP, Brazil
                [6 ]Department of Implantology, University Santo Amaro – UNISA, São Paulo, SP, Brazil
                Author notes
                *Author for correspondence: vanessa.penn@ 123456gmail.com
                Article
                10.4155/fso.15.26
                5137914
                eca4806d-2539-44d0-a36a-38ef832d3e4e
                © Vanessa Penna et al.

                This work is licensed under a Creative Commons Attribution 4.0 License

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                cell communication,cell culture,extracellular matrix,mechanotransduction,proteolytic enzymes,regenerative medicine,stem cells,surface proteins,tissue engineering

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