Investigating The Development Of Midbrain Dopaminergic Neurons Using Mouse Embryonic Stem Cell Reporter Lines

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Investigating the Development of Midbrain Dopaminergic Neurons Using Mouse Embryonic Stem Cell Reporter Lines

Author : Colin Tze En Su
Publisher :
ISBN 13 :
Total Pages : 490 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis Investigating the Development of Midbrain Dopaminergic Neurons Using Mouse Embryonic Stem Cell Reporter Lines by : Colin Tze En Su

Download or read book Investigating the Development of Midbrain Dopaminergic Neurons Using Mouse Embryonic Stem Cell Reporter Lines written by Colin Tze En Su and published by . This book was released on 2012 with total page 490 pages. Available in PDF, EPUB and Kindle. Book excerpt: Embryonic stem (ES) cells possess the capability to self-renew indefinitely and are capable of generating any cell of the three primary germ layers, making them an attractive source of material to investigate both basic physiological properties and neurodegenerative processes. Although ES cells can be directed into specific cell lineages, the differentiation of ES cells results in heterogenous cultures. To date, there are few differentiation protocols that produce homogenous populations of any desired cell type. Many methods have been used in an effort to obtain homogenous populations of cells; from forced expression of genes involved in developmental pathways, to FACS isolation of cells expressing markers of interest. There has been a considerable focus on generating homogenous populations of midbrain dopaminergic progenitors (or neurons) for Parkinson's disease which involves the degeneration of a specific population of midbrain dopaminergic neurons. In this thesis, I investigate the development of mouse embryonic stem cells into midbrain dopaminergic neurons using reporter cell lines. In the first experimental chapter, I investigate the expression of Lmx1a and Msx1; two key transcription factors implicated in dopaminergic neuronal development. I also examine the impact of the BMP, Shh and Wnt signalling pathways on dopaminergic neural differentiation. Activation of the BMP and Wnt pathways resulted in inhibition of neural induction and the expression of both Lmx1a and Msx1. In contrast, antagonising these signalling pathways increased the yield of tyrosine hydroxylase (TH) expressing neurons. Activating or inhibiting the Shh pathway did not affect Lmx1a, Msx1 or TH expression. These experiments show that early Lmx1a expression is not indicative of the number of dopaminergic neurons produced. Furthermore, many of the TH positive neurons derived from monolayer cultures were not of midbrain origin. In the following experimental chapter, I used immunocytochemistry and qPCR to characterise the population of cells expressing Lmx1a. The downstream targets of Lmx1a, Msx1 and Wnt1, and midbrain dopaminergic neuron markers, Lmx1b and En1, were significantly upregulated in Lmx1a positive cells. The Lmx1a positive fraction was enriched with neural progenitors, and give rise to highly neural cultures. However, the majority of neurons in the terminally differentiated cultures derived from Lmx1a positive cells were GABAergic. Immunocytochemistry identified these cells as forebrain GABAergic neurons with upper-layer identity. Furthermore, the isolated Lmx1a positive cells were not responsive to patterning cues, indicating that they were already committed towards a GABAergic neuron fate. To show that these Lmx1a+ progenitors could generate dopaminergic neurons I used an alternative differentiation paradigm, the PA6 co-culture method. Expression of Lmx1a in PA6 co-cultures was different from monolayer cultures; the percentage of Lmx1a positive cells increased throughout the differentiation period. In addition, PA6 co-culture derived TH positive cells were found to co-express Lmx1a, an occurrence that was uncommon in monolayer cultures.The ionotropic glutamate receptors on neurons derived on adherent monolayer and PA6 co-cultures were functionally characterised in the final experimental chapter. Previously, antagonism of ionotropic glutamate receptors has been reported to improve behavioural assay scores in Parkinsonian animal models (Johnson et al., 2009). Terminally differentiated monolayer cultures and PA6 co-cultures responded differently to stimulation with glutamate, AMPA kainate and NMDA. The ionotropic glutamate receptors of midbrain dopaminergic and GABAergic neurons derived from both culture systems were further investigated. An initial characterisation indicates distinct differences between the glutamate receptor populations in monolayer and PA6 co-cultures. It appears that monolayer differentiation generates AMPA expressing midbrain dopaminergic neurons in comparison to the NMDA receptors evident following PA6 differentiation. Interestingly, these differences in receptor expression appear restricted by culture method, rather than neuronal subtype, i.e. monolayer neurons expressed AMPA receptors, regardless of whether they were TH+ or GAD67+. Similarly both TH+ and GAD67+ neurons appeared to express NMDA receptors following PA6 differentiation. At present the significance of these findings is unknown. In addition, the effect of Wnt5a on cell responses to glutamate agonists was examined. Wnt5a was able to potentiate cell responses to sub-maximal concentrations of certain glutamate agonists depending on the differentiation paradigm performed.

Generation of Mouse Embryonic Stem Cell Reporter Lines to Identify Developmental Stages During Induction of Dopaminergic Neurons

Author : Rhys Henry Stone Bellinge
Publisher :
ISBN 13 :
Total Pages : 151 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis Generation of Mouse Embryonic Stem Cell Reporter Lines to Identify Developmental Stages During Induction of Dopaminergic Neurons by : Rhys Henry Stone Bellinge

Download or read book Generation of Mouse Embryonic Stem Cell Reporter Lines to Identify Developmental Stages During Induction of Dopaminergic Neurons written by Rhys Henry Stone Bellinge and published by . This book was released on 2013 with total page 151 pages. Available in PDF, EPUB and Kindle. Book excerpt: The most fundamental questions on how neurons arise from specific developmental programs focus on how they acquire their correct synaptic connections, and how they mature and develop their distinct phenotypes. It is known in a broad sense that each group of neurons in the functional circuits of the adult brain develops according to both intrinsic programming of molecular cascades, and extrinsic influences from their environments. Here I set out to investigate the cells of the rostral region of the central nervous system, those of themesencephalon, or midbrain, by the use of murine embryonic stem (ES) cells as an in vitro model of midbrain dopaminergic (mDA) progenitor cells. The focus of the work was to introduce genetic reporters into ES cells to allow identification and potentially isolation of neuronal progenitors during in vitro differentiation of ES cells. mDA neurons arise in the ventral midline of the midbrain, and are intrinsic in such functions as fine motor control, emotion, perception and higher cognition, as well as learning and reward. Loss of these neurons in the substantia nigra leads to the symptoms of Parkinson's disease (PD). Before ES cell therapies can attempt to treat PD, the signalling factors involved in the early specification of mDA neurons need to be thoroughly characterised. Differentiation of ES cells using existing protocols does not result in homogeneous populations of neural progenitors. Typically, mouse ES cells differentiate into neural progenitors by default once ES cell growth supplements are removed, but rarely are these mDA progenitor neurons. In this thesis, the neural induction protocol was first investigated to examine whether it was generating neural progenitors of a midbrain phenotype using an Lmx1a-eGFP reporter cell line, as a study control. The induction protocol was found to generate, at its peak, 8.1% of the total cell population expressing the fluorescent reporter. This indicated that the induction protocol was not yielding enriched populations of Lmx1a+ neural progenitors. Additionally it was considered that it may be that not all of the Lmx1a-eGFP+ cells were destined to become dopaminergic neurons, and that perhaps more reporters were needed to identify midbraindopaminergic neurons. Following this, a series of homologous recombination experiments were performed, initially successfully producing an Lmx1b-CBR-IRES-dsRed reporter in a murine ES (mES) cell line, with 17.9% targeting efficiency. Following this a series of targeting experiments were carried out in an attempt to target a Pitx3-mCitrine reporter to an existing Lmx1a-AMP mES reporter cell line. Unfortunately, despite the expansion and screening of over 1000 antibiotic resistant colonies over a 15 month period, this targeting campaign was not successful. In an effort to assess the cause for this lack of success, an additional targeting experiment was successfully performed, using an Lmx1a-mCFP targeting vector to recombine with a wild-type mES cell line, with a 3.45% targeting efficiency. In all likelihood the failure of the Pitx3-mCitrine to target adequately lies with the design of the targeting vector and primers used; the homology of the vector to host genome was 7kb in total, which may in fact have been in excess when considering that the target, Pitx3, is expressed late in the differentiation of DA neurons, giving rise to the possibility that the DNA is tightly bound and comparatively inaccessible at this undifferentiated state. The intention for the generation of dual Lmx1a-postive/Pitx3-positive fluorescent reporter cell lines was to enable the identification of DA neuronal progenitors, with the goal of ultimately using these cells to isolate DA cells at definite specific stages of development prior to transplantation into a mouse model of PD. Although the Pitx3-mCitrine targeting did not succeed, the neural induction protocol was successfully assayed using an Lmx1a-eGFP reporter cell line, a novel Lmx1a-mCFP reporter cell line was created, as was a novel Lmx1b-CBR-dsRed cell line.

Regulating Dopaminergic Development of Mouse Embryonic Stem Cells Using Small Molecules

Author : Cameron Philip James Hunt
Publisher :
ISBN 13 :
Total Pages : 732 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis Regulating Dopaminergic Development of Mouse Embryonic Stem Cells Using Small Molecules by : Cameron Philip James Hunt

Download or read book Regulating Dopaminergic Development of Mouse Embryonic Stem Cells Using Small Molecules written by Cameron Philip James Hunt and published by . This book was released on 2013 with total page 732 pages. Available in PDF, EPUB and Kindle. Book excerpt: The degeneration of substantia nigral dopaminergic (mDA) neurons is responsible for the resting tremors and difficulties in fine motor control seen in Parkinson's disease. Embryonic stem cell (ESC)-derived mDA neurons provide an ideal opportunity to model mDA development in vitro. However, current mouse differentiation protocols are inefficient possibly as a result of the influence of endogenous signalling pathways. In theory, the ability to identify cell types that have undergone specification of neural lineage should make it easier to identify upstream regulators of that lineage. In this study dopaminergic progenitor specific reporter cell lines expressing -lactamase or luciferase under the control of the Lmx1a promoter were used to investigate protein kinase signalling pathways involved in dopaminergic neural specification.Screening experiments using a small kinase inhibitor library revealed that inhibition of Epidermal Growth Factor (EGF), Vascular Endothelial Growth Factor (VEGF) and DNA-dependent protein kinase (DNA-PK) signalling pathways promoted Lmx1a expression using Lmx1a reporter mESC lines. Moreover, additional experiments revealed that DNA-PKi-mediated increases in Lmx1a+ progenitors upregulated Notch signalling suggesting that DNA-PK signalling may alter neurogenic potential of developing progenitors. Co-culture differentiation revealed that DNA-PKi alone positively regulated Lmx1a expression. However, as the generation of Nurr1+ post-mitotic dopaminergic progenitors could not be enhanced using small molecules identified from early screens suggests that these compounds may exclusively regulate earlier specification pathways such as Lmx1a. Intriguingly, only progenitors with compromised EGF signalling were able promote generation of dopaminergic (TH+) neurons.Lastly, inhibition of endogenous Smad signalling pathways sped up neural induction and promoted lineage specification within mESCs, albeit within a narrow time frame in a monolayer. Specific inhibition of the Smad1/5/8 pathway induced a floor plate (FP) transcription factor profile with an upregulation of Corin, Shh and FoxA2 genes and increased the number of TH+ neurons. The FP molecular signature was further enhanced by the addition of small molecules CHIR and SAG that promote Wnt and Shh signalling pathways, respectively. Although these studies demonstrated that midbrain and FP specification could be regulated in mESCs using inhibitors for DNA-PK and Smad signalling pathways, co-incubation with DNA-PK and Smad inhibitors did not support ventral mDA specification. This may be due to the immature or neurogenic state generated by DNA-PK inhibition to support neural specification driven by inhibition of Smad pathways. These studies, demonstrate that DNA-PK signalling has a, previously unrecognised, role in regulating neural specification and that endogenous Smad signalling can rapidly specify neuroectoderm and regional subtype specification in a monolayer. These studies provide insight into the complex regulation of endogenous signalling networks during neural differentiation of mESCs and how they may be regulated using targeted small molecules.

Defining Conditions Required for the Derivation of Midbrain Dopaminergic Neurons from Embryonic Stem Cells Using Stem Cell Reporter Lines

Download Defining Conditions Required for the Derivation of Midbrain Dopaminergic Neurons from Embryonic Stem Cells Using Stem Cell Reporter Lines PDF Online Free

Author : Brigham Jay Hartley
Publisher :
ISBN 13 :
Total Pages : 566 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis Defining Conditions Required for the Derivation of Midbrain Dopaminergic Neurons from Embryonic Stem Cells Using Stem Cell Reporter Lines by : Brigham Jay Hartley

Download or read book Defining Conditions Required for the Derivation of Midbrain Dopaminergic Neurons from Embryonic Stem Cells Using Stem Cell Reporter Lines written by Brigham Jay Hartley and published by . This book was released on 2013 with total page 566 pages. Available in PDF, EPUB and Kindle. Book excerpt: The efficient and robust derivation of midbrain dopaminergic (mbDA) cells from embryonic stem cells (ESCs) may pave the way for cell replacement therapy (CRT) for Parkinson's Disease (PD) patients whose disease is refractory to current pharmacotherapy. It may also facilitate investigation of the molecular mechanisms underlying the pathogenesis of PD through in vitro modeling. However, a major hindrance to the application of ESC-derived cells for in vitro and in vivo applications is the observation that current differentiation protocols yield heterogeneous cultures. Reporter cell lines offer the ability to isolate cells from differentiating cultures. The LIM homeobox transcription factor 1 [alpha] (LMX1A) is a prominent regulator of early mbDA and forebrain GABAergic specification. In this thesis I utilize both mouse and human ESCs expressing EGFP under the control of the endogenous LMX1A promoter to investigate aspects of fate specification following derivation under a chemically defined monolayer (CDML) or PA6 stromal cell differentiation protocols. ESC differentiation under PA6 conditions results in a population of mbDA progenitors that can be isolated based on EGFP expression, which subsequently gives rise to high yields of mbDA neurons. ESCs differentiating under CDML neural induction cues only, default to an anterior neuroectoderm phenotype. However, timed exposure to morphogens that mimic in vivo mbDA developmental conditions (SHH & WNT signalling) derives functional DA neurons with a protein profile indicative of a midbrain phenotype. The cultivation and subsequent subtype specification of these defined neural progenitors holds promise for the derivation of mbDA cells for PD CRT, in vitro disease modelling and pharmacotherapeutic screening platforms.

Differentiation of Embryonic Stem Cells

Author :
Publisher : Elsevier
ISBN 13 : 0080546161
Total Pages : 577 pages
Book Rating : 4.0/5 (85 download)

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Book Synopsis Differentiation of Embryonic Stem Cells by :

Download or read book Differentiation of Embryonic Stem Cells written by and published by Elsevier. This book was released on 2003-12-18 with total page 577 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume covers all aspects of embryonic stem cell differentiation, including mouse embryonic stem cells, mouse embryonic germ cells, monkey and human embryonic stem cells, and gene discovery. * Early commitment steps and generation of chimeric mice* Differentiation to mesoderm derivatives* Gene discovery by manipulation of mouse embryonic stem cells

Stem Cells in Reproductive Medicine

Author : Carlos Simón
Publisher : Cambridge University Press
ISBN 13 : 1107034477
Total Pages : 199 pages
Book Rating : 4.1/5 (7 download)

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Book Synopsis Stem Cells in Reproductive Medicine by : Carlos Simón

Download or read book Stem Cells in Reproductive Medicine written by Carlos Simón and published by Cambridge University Press. This book was released on 2013-07-04 with total page 199 pages. Available in PDF, EPUB and Kindle. Book excerpt: Stem cell science has the potential to impact human reproductive medicine significantly - cutting edge technologies allow the production and regeneration of viable gametes from human stem cells offering potential to preciously infertile patients. Written by leading experts in the field Stem Cells in Reproductive Medicine brings together chapters on the genetics and epigenetics of both the male and female gametes as well as advice on the production and regeneration of gene cells in men and women, trophoblasts and endometrium from human embryonic and adult stem cells. Although focussing mainly on the practical elements of the use of stem cells in reproductive medicine, the book also contains a section on new developments in stem cell research. The book is essential reading for reproductive medicine clinicians, gynecologists and embryologists who want to keep abreast of practical developments in this rapidly developing field.

Human Neural Stem Cells

Author : Leonora Buzanska
Publisher : Springer
ISBN 13 : 3319934856
Total Pages : 329 pages
Book Rating : 4.3/5 (199 download)

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Book Synopsis Human Neural Stem Cells by : Leonora Buzanska

Download or read book Human Neural Stem Cells written by Leonora Buzanska and published by Springer. This book was released on 2018-09-12 with total page 329 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book summarizes early pioneering achievements in the field of human neural stem cell (hNSC) research and combines them with the latest advances in stem cell technology, including reprogramming and gene editing. The powerful potential of hNSC to generate and repair the developing and adult CNS has been confirmed by numerous experimental in vitro and in vivo studies. The book presents methods for hNSC derivation and discusses the mechanisms underlying NSC in vitro fate decisions and their in vivo therapeutic mode of action. The long-standing dogma that the human central nervous system (CNS) lacks the ability to regenerate was refuted at the end of the 20th century, when evidence of the presence of neurogenic zones in the adult human brain was found. These neurogenic zones are home to human neural stem cells (hNSCs), which are capable of self-renewing and differentiating into neurons, astrocytes and oligodendrocytes. NSCs isolated from human CNS have a number of clinical advantages, especially the innate potential to differentiate into functional neural cells. Nevertheless, their full clinical exploitation has been hindered by limited access to the tissue and low expansion potential. The search for an alternative to CNS sources of autologous, therapeutically competent hNSCs was the driving force for the many studies proving the in vitro plasticity of different somatic stem cells to generate NSCs and their functional progeny. Now the era of induced pluripotent stem cells has opened entirely new opportunities to achieve research and therapeutic goals with the aid of hNSCs.

Mouse Brain Development

Author : Andre M. Goffinet
Publisher : Springer Science & Business Media
ISBN 13 : 3540480021
Total Pages : 347 pages
Book Rating : 4.5/5 (44 download)

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Book Synopsis Mouse Brain Development by : Andre M. Goffinet

Download or read book Mouse Brain Development written by Andre M. Goffinet and published by Springer Science & Business Media. This book was released on 2012-08-10 with total page 347 pages. Available in PDF, EPUB and Kindle. Book excerpt: Our understanding of the molecular mechanisms involved in mammalian brain development remains limited. However, the last few years have wit nessed a quantum leap in our knowledge, due to technological improve ments, particularly in molecular genetics. Despite this progress, the available body of data remains mostly phenomenological and reveals very little about the grammar that organizes the molecular dictionary to articulate a pheno type. Nevertheless, the recent progress in genetics will allow us to contem plate, for the first time, the integration of observation into a coherent view of brain development. Clearly, this may be a major challenge for the next century, and arguably is the most important task of contemporary develop mental biology. The purpose of the present book is to provide an overview that syn thesizes up-to-date information on selected aspects of mouse brain devel opment. Given the format, it was not possible to cover all aspects of brain development, and many important subjects are missing. The selected themes are, to a certain extent, subjective and reflect the interests of the contributing authors. Examples of major themes that are not covered are peripheral nervous system development, including myelination, the development of the hippocampus and several other CNS structures, as well as the developmental function of some important morphoregulatory molecules.

In Situ Hybridization

Author : D. G. Wilkinson
Publisher : Oxford University Press, USA
ISBN 13 : 9780199636587
Total Pages : 224 pages
Book Rating : 4.6/5 (365 download)

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Book Synopsis In Situ Hybridization by : D. G. Wilkinson

Download or read book In Situ Hybridization written by D. G. Wilkinson and published by Oxford University Press, USA. This book was released on 1998 with total page 224 pages. Available in PDF, EPUB and Kindle. Book excerpt: In situ hybridization is used to reveal the location of specific nucleic acids sequences on chromosomes or in tissues. Visualization of the location of genes on chromosomes or of specific mRNAs or viruses in tissues is crucial for understanding the organization, regulation, and function of genes. It is a therefore a core technique in all areas of biomedical research. In Situ Hybridization: A Practical Approach 2/e is the second edition of one of the most successful Practical Approach books, published in 1992. Since the first edition was published, a number of important technical advances have been made. The new edition has been thoroughly updated to contain protocols detailing the major techniques of in situ hybridization currently in use: in situ hybridization to mRNA with oligonucleotide and RNA probes (radiolabelled and hapten labelled); analysis using light and electron microscopes; whole mount in situ hybridization; double detection of RNAs, and RNA plus protein; and fluorescent in situ hybridization to detect chromosomal sequences. The protocols are complemented by advice on strategies for successful results, descriptions of the theoretical basis of in situ hybridization and important new developments in gene expression databases. The procedures described are widely applicable to many systems. The use of in situ hybridization in PCR is covered in a separate volume: Herrington and O'Leary (Eds) PCR 3 - PCR in situ hybridization: A Practical Approach (OUP, 1997). All the authors have extensive practical experience of establishing reliable techniques of in situ hybridization. This book will be useful to all researchers at all levels who use in situ hybridization.

Human Embryonic Stem Cells

Author : Arlene Chiu
Publisher : Springer Science & Business Media
ISBN 13 :
Total Pages : 488 pages
Book Rating : 4.3/5 (91 download)

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Book Synopsis Human Embryonic Stem Cells by : Arlene Chiu

Download or read book Human Embryonic Stem Cells written by Arlene Chiu and published by Springer Science & Business Media. This book was released on 2003-08 with total page 488 pages. Available in PDF, EPUB and Kindle. Book excerpt: A discussion of all the key issues in the use of human pluripotent stem cells for treating degenerative diseases or for replacing tissues lost from trauma. On the practical side, the topics range from the problems of deriving human embryonic stem cells and driving their differentiation along specific lineages, regulating their development into mature cells, and bringing stem cell therapy to clinical trials. Regulatory issues are addressed in discussions of the ethical debate surrounding the derivation of human embryonic stem cells and the current policies governing their use in the United States and abroad, including the rules and conditions regulating federal funding and questions of intellectual property.

Regenerative Pharmacology

Author : George J. Christ
Publisher : Cambridge University Press
ISBN 13 : 0521899494
Total Pages : 361 pages
Book Rating : 4.5/5 (218 download)

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Book Synopsis Regenerative Pharmacology by : George J. Christ

Download or read book Regenerative Pharmacology written by George J. Christ and published by Cambridge University Press. This book was released on 2013-04-15 with total page 361 pages. Available in PDF, EPUB and Kindle. Book excerpt: A state-of-the-art primer on the role of pharmacological sciences in regenerative medicine, for advanced students, postdoctoral fellows, and researchers.

Stem Cells

Author : Ariff Bongso
Publisher : World Scientific
ISBN 13 : 9814289388
Total Pages : 721 pages
Book Rating : 4.8/5 (142 download)

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Book Synopsis Stem Cells by : Ariff Bongso

Download or read book Stem Cells written by Ariff Bongso and published by World Scientific. This book was released on 2011 with total page 721 pages. Available in PDF, EPUB and Kindle. Book excerpt: Stem cell biology has drawn tremendous interest in recent years as it promises cures for a variety of incurable diseases. This book deals with the basic and clinical aspects of stem cell research and involves work on the full spectrum of stem cells isolated today. It also covers the conversion of stem cell types into a variety of useful tissues which may be used in the future for transplantation therapy. It is thus aimed at undergraduates, postgraduates, scientists, embryologists, doctors, tissue engineers and anyone who wishes to gain some insight into stem cell biology. This book is important as it is comprehensive and covers all aspects of stem cell biology, from basic research to clinical applications. It will have 33 chapters written by renowned stem cell scientists worldwide. It will be up-to-date and all the chapters include self-explanatory figures, color photographs, graphics and tables. It will be easy to read and give the reader a complete understanding and state of the art of the exciting science and its applications.

Cell Engineering and Regeneration

Author : Heinz Redl
Publisher : Springer
ISBN 13 : 9783319088303
Total Pages : 0 pages
Book Rating : 4.0/5 (883 download)

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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 and published by Springer. This book was released on 2017-02-16 with total page 0 pages. 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.

Quantitative Methods in Neuroscience

Author : Stephen M. Evans
Publisher :
ISBN 13 : 9780198505280
Total Pages : 362 pages
Book Rating : 4.5/5 (52 download)

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Book Synopsis Quantitative Methods in Neuroscience by : Stephen M. Evans

Download or read book Quantitative Methods in Neuroscience written by Stephen M. Evans and published by . This book was released on 2004 with total page 362 pages. Available in PDF, EPUB and Kindle. Book excerpt: Stereology is a valuable tool for neuroscientists, allowing them to obtain 3-Dimensional information from 2-Dimensional measurements made on appropriately sampled sections (usually obtained from histological sections or MRI/CT/PET scans). This 3-D information is invaluable in correlatingstructural/functional relationships in the pursuit of far greater understanding of the function of the central nervous system. However, in carrying out such measurements, often based on limited data sets, there is a risk of experimenter bias. An important feature of modern design based stereology isto be aware of potential sources of bias and eliminate them during the data collection. With many of the major neuroscience journals now insisting that quantitative data be presented, there is a greater need than ever for neuroscientists to understand the theory and practice behind quantitativemethods, such as those offered by stereology. Quantitative Methods in Neuroscience is a cookbook of stereological methods written especially for neuroscientists. It provides clear and accessible advice about when and when not to use stereology. Throughout the book, the emphasis is on practical guidance, rather than discussions and formulae.Written by leading scientists in the field of stereology, with a Foreword by D.C. Sterio, the book will be a valuable introduction to these methods for neuroscientists, and all those involved in development of new drug programmes.

Human Stem Cell Manual

Author : Suzanne Peterson
Publisher : Academic Press
ISBN 13 : 0123854741
Total Pages : 652 pages
Book Rating : 4.1/5 (238 download)

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Book Synopsis Human Stem Cell Manual by : Suzanne Peterson

Download or read book Human Stem Cell Manual written by Suzanne Peterson and published by Academic Press. This book was released on 2012-10-22 with total page 652 pages. Available in PDF, EPUB and Kindle. Book excerpt: This manual is a comprehensive compilation of "methods that work" for deriving, characterizing, and differentiating hPSCs, written by the researchers who developed and tested the methods and use them every day in their laboratories. The manual is much more than a collection of recipes; it is intended to spark the interest of scientists in areas of stem cell biology that they may not have considered to be important to their work. The second edition of the Human Stem Cell Manual is an extraordinary laboratory guide for both experienced stem cell researchers and those just beginning to use stem cells in their work. Offers a comprehensive guide for medical and biology researchers who want to use stem cells for basic research, disease modeling, drug development, and cell therapy applications Provides a cohesive global view of the current state of stem cell research, with chapters written by pioneering stem cell researchers in Asia, Europe, and North America Includes new chapters devoted to recently developed methods, such as iPSC technology, written by the scientists who made these breakthroughs

Stem Cell Biology

Author : Daniel R. Marshak
Publisher : CSHL Press
ISBN 13 : 9780879696733
Total Pages : 566 pages
Book Rating : 4.6/5 (967 download)

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Book Synopsis Stem Cell Biology by : Daniel R. Marshak

Download or read book Stem Cell Biology written by Daniel R. Marshak and published by CSHL Press. This book was released on 2001 with total page 566 pages. Available in PDF, EPUB and Kindle. Book excerpt: Stem cells are the focus of intense interest from a growing, multidisciplinary community of investigators with new tools for isolating and characterizing these elusive cell types. This volume, which features contributions from many of the world's leading laboratories, provides a uniquely broad and authoritative basis for understanding the biology of stem cells and the current excitement about their potential for clinical exploitation. It is an essential work of reference for investigators in embryology, hematology, and neurobiology, and their potential for clinical exploitation. It is an essential work of reference for investigators in embryology, hematology, and neurobiology, and their collaborators in the emerging field of regenerative medicine.

Neural Crest Cells

Author : Quenten Schwarz
Publisher : Humana Press
ISBN 13 : 9781493994113
Total Pages : 230 pages
Book Rating : 4.9/5 (941 download)

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Book Synopsis Neural Crest Cells by : Quenten Schwarz

Download or read book Neural Crest Cells written by Quenten Schwarz and published by Humana Press. This book was released on 2019-06-06 with total page 230 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed book provides a wide range of techniques, from those that have been used extensively since the very first investigations into neural crest cells to those that are currently cutting-edge, in order to explore the development of neural crest cells in human, mice, rat, chick, quail, medaka, and shark. With a bit of imagination and adjustment, many of these methodologies can be adaptable to any species desirable for study. Written for the highly successful Methods in Molecular Biology series format, 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. Authoritative and practical, Neural Crest Cells: Methods and Protocols serves as an ideal reference guide for aspiring and experienced developmental biologists alike.