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Expansion of Human Induced Pluripotent Stem Cells on Synthetic Substrate in Defined Medium

2012 Stem cells
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Release : 2012
Genre : Stem cells
Kind : eBook
Book Rating : 349/5 ( reviews)

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Book Synopsis Expansion of Human Induced Pluripotent Stem Cells on Synthetic Substrate in Defined Medium by : Huantong Yao

Download or read book Expansion of Human Induced Pluripotent Stem Cells on Synthetic Substrate in Defined Medium written by Huantong Yao. This book was released on 2012. Available in PDF, EPUB and Kindle. Book excerpt: Human induced pluripotent stem cells (hiPSCs) have the potential to generate patient-specific cells to treat many incurable diseases by cell replacement therapy. However, so far the culture of hiPSCs depends greatly on feeder cells or Matrigel which has safety issues. Thus, chemically defined substrates that could provide niches necessary for cell attachment and proliferation are preferred for clinical application of hiPSCs. Recently, Corning Life Sciences has developed synthetic peptide-functionalized cell culture surface, referred to as Corning RTM Synthemax(TM) that support self-renewal and differentiation of human embryonic stem cell (hESC). In this work, we have collaborated with Corning to investigate the attachment, proliferation, and differentiation of hiPSCs on the Synthemax substrate. We demonstrated that iPS cells retained stable proliferation and pluripotency marker protein expression after growing on the Synthemax substrate for ten consecutive passages. Further examination reveals that integrins alphaVbeta5 mediates attachment to the substrate. Moreover, we observed hiPSCs colonies were more compact on the Synthemax surface. This may be due to less activation of beta-catenin-mediated Wnt signaling pathway in cells on the synthetic peptide surface. In hiPSCs grown on the Synthemax Surface, we also found denser actin filaments in the cell-cell interface and down-regulation of vinculin and up-regulation of zyxin, indicating the reorganization of cytoskeleton structure inside cells in response to cell-matrix interaction.

Stem Cell Engineering

2012-11-20 Medical
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Release : 2012-11-20
Genre : Medical
Kind : eBook
Book Rating : 04X/5 ( reviews)

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Book Synopsis Stem Cell Engineering by : David Schaffer

Download or read book Stem Cell Engineering written by David Schaffer. This book was released on 2012-11-20. Available in PDF, EPUB and Kindle. Book excerpt: While the potential of stem cells is recognized, their proliferation and differentiation must be more precisely controlled to maximize the production of therapeutically relevant cells and for cell replacement therapies to minimize contamination with residual cells that can give rise to side effects. How can engineers make contributions to address these challenges? With contributions from pioneers and experts, Stem Cell Engineering: Principles and Practices highlights recent advances in the understanding of the cellular and molecular composition of the stem cell niche, as well as approaches to build upon this basic information to direct stem cell differentiation into therapeutically valuable lineages. The growing recognition of stem cells as an important and exciting field will continue to draw investigators with diverse backgrounds—from biology, engineering, and the physical sciences—and thereby enable further progress in these and other new directions. This book discusses advances made during the last decade that have led to increasingly defined culture systems for growing stem cells, starting from co-culture with feeder cells in the presence of serum to growth on synthetic substrates in defined medium. In addition to highlighting many recent advances, it underscores the need for future work.

Induced Pluripotent Stem (iPS) Cells

2022-06-22 Science
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Release : 2022-06-22
Genre : Science
Kind : eBook
Book Rating : 19X/5 ( reviews)

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Book Synopsis Induced Pluripotent Stem (iPS) Cells by : Andras Nagy

Download or read book Induced Pluripotent Stem (iPS) Cells written by Andras Nagy. This book was released on 2022-06-22. Available in PDF, EPUB and Kindle. Book excerpt: This extensive new edition presents protocols reflecting the great strides made in the study of induced pluripotent stem (iPS) cells. The collection explores new and improved methods for the generation, expansion, and maintenance of iPS cells from different tissue types, characterization of their differentiation pathways along different lineages, and their potential utility in tissue repair and regeneration. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Induced Pluripotent Stem (iPS) Cells: Methods and Protocols, Second Edition aims to arm stem cell biologists, both novice and expert, with invaluable protocols that are currently being used in various laboratories around the world.

Expansion of Human Pluripotent Stem Cells with Synthetic Oolymer PMVE-alt-MA

2011
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Release : 2011
Genre :
Kind : eBook
Book Rating : 305/5 ( reviews)

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Book Synopsis Expansion of Human Pluripotent Stem Cells with Synthetic Oolymer PMVE-alt-MA by : Wai Keung Chu

Download or read book Expansion of Human Pluripotent Stem Cells with Synthetic Oolymer PMVE-alt-MA written by Wai Keung Chu. This book was released on 2011. Available in PDF, EPUB and Kindle. Book excerpt: The differentiation potential of human pluripotent stem cells (hPSCs) promises to treat degenerative diseases with cell replacement therapies. Current culture media and substrates are not only expensive, but are undefined for clinical use because of the various known and unknown animal components that are required to promote proliferation and differentiation. Chemically defined materials are ideal substitutes for hPSC culture. This study focused on the use of PMVE-alt-MA, a synthetic polymer that was previously identified by arrayed screening technology for ex vivo hPSCs maintenance. Synthetic hydrogels and microcarriers endowed with PMVE-alt-MA moieties were used to evaluate hPSC expansion, adhesion, proliferation, colony formation and maintenance. Flow cytometry and realtime PCR were used to characterize the expression of pluripotency markers in both hydrogel and microcarrier cultures. Protein adsorption to PMVE-alt-MA was found necessary for initial cell adhesion. Enzymatic dissociation with Accutase TM was optimized to passage the embryonic stem cell line Hues9 on 9.7% semi-IPN hydrogel for 8 passages and the result was comparable to Matrigel culture. However, polymer coatings on polyacrylamide hydrogel and microcarriers were found to be insufficient for stem cell self-renewal and attachment respectively. Embryoid bodies (EBs) with uniform size and shape were observed in microcarrier suspension but no difference was found in the differentiation pattern when compared to normal suspension EB formation. Our experimental results demonstrated the potential to use PMVE-alt-MA in hPSC expansion, as expected from the arrayed screening technology.

Cell Engineering and Regeneration

2017-02-16 Technology & Engineering
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Release : 2017-02-16
Genre : Technology & Engineering
Kind : eBook
Book Rating : 303/5 ( reviews)

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Book Synopsis Cell Engineering and Regeneration by : Heinz Redl

Download or read book Cell Engineering and Regeneration written by Heinz Redl. This book was released on 2017-02-16. Available in PDF, EPUB and Kindle. Book excerpt: This reference work presents the origins of cells for tissue engineering and regeneration, including primary cells, tissue-specific stem cells, pluripotent stem cells and trans-differentiated or reprogrammed cells. There is particular emphasis on current understanding of tissue regeneration based on embryology and evolution studies, including mechanisms of amphibian regeneration. The book covers the use of autologous versus allogeneic cell sources, as well as various procedures used for cell isolation and cell pre-conditioning , such as cell sorting, biochemical and biophysical pre-conditioning, transfection and aggregation. It also presents cell modulation using growth factors, molecular factors, epigenetic approaches, changes in biophysical environment, cellular co-culture and other elements of the cellular microenvironment. The pathways of cell delivery are discussed with respect to specific clinical situations, including delivery of ex vivo manipulated cells via local and systemic routes, as well as activation and migration of endogenous reservoirs of reparative cells. The volume concludes with an in-depth discussion of the tracking of cells in vivo and their various regenerative activities inside the body, including differentiation, new tissue formation and actions on other cells by direct cell-to-cell communication and by secretion of biomolecules.

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