Tissue Culture Of The Nervous System

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Tissue Culture of the Nervous System

Author : Gordon Sato
Publisher : Springer Science & Business Media
ISBN 13 : 1468429043
Total Pages : 298 pages
Book Rating : 4.4/5 (684 download)

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Book Synopsis Tissue Culture of the Nervous System by : Gordon Sato

Download or read book Tissue Culture of the Nervous System written by Gordon Sato and published by Springer Science & Business Media. This book was released on 2013-06-29 with total page 298 pages. Available in PDF, EPUB and Kindle. Book excerpt: The impetus for compiling this book was the recent development of culture strains of neuroblastoma and glial cells and the immediate and enthusiastic way they have been taken up as model systems. After the first sudden rush of activity, it seems appropriate to pause, to assess progress, and to contemplate the future contributions that may be possible using these culture techniques. Long before the advent of established strains, cultures of nervous tissue had already contributed to neurobiology. Ross Harrison, in 1906, in a single experimental series, established tissue culture as a promising new technique in cell biology and settled the Golgi-Cajal controversy as to whether axonic processes originated as outgrowths from the cell body or were formed first in the intercellular spaces and were later connected to the cell body. Harrison observed process growth from nerve cells in cultures, thus settling the matter in favor of Cajal. Of great importance to neurobiology is the discovery by Rita Levi-Montalcini of nerve growth factor. Cultures of spinal ganglia played a major role in the discovery, isolation, and characterization of the factor (Levi-Montalcini et ai. , 1954). In my opinion, this discovery, although very well known, has not yet been adequately recognized for its germinal influence on neurobiology and embryology. Progress since the advent of clonal cultures has been more modest. I would like to cite two pieces of work which emphasize the technical ad vantages of these cultures.

Neuronal Cell Culture

Author : Shohreh Amini
Publisher : Humana
ISBN 13 : 9781071614396
Total Pages : 205 pages
Book Rating : 4.6/5 (143 download)

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Book Synopsis Neuronal Cell Culture by : Shohreh Amini

Download or read book Neuronal Cell Culture written by Shohreh Amini and published by Humana. This book was released on 2022-06-09 with total page 205 pages. Available in PDF, EPUB and Kindle. Book excerpt: This second edition volume details the latest aspects of neural cells covering the practical and theoretical considerations of each techniques involved. Chapters guide readers through a general overview of the neuronal culturing principles, cell line models for neural cells, the isolation and propagation of primary cultures, stem cells, transfection and transduction of neural cultures, and other more advanced techniques. Written in 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. Practical and easy to use, Neuronal Cell Culture: Methods and Protocols, Second Edition aims to be of interest to scientists at all levels studying cell culture models for neuroscientific studies.

A Dissection and Tissue Culture Manual of the Nervous System

Author : Abraham Shahar
Publisher : Wiley-Liss
ISBN 13 :
Total Pages : 408 pages
Book Rating : 4.3/5 (91 download)

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Book Synopsis A Dissection and Tissue Culture Manual of the Nervous System by : Abraham Shahar

Download or read book A Dissection and Tissue Culture Manual of the Nervous System written by Abraham Shahar and published by Wiley-Liss. This book was released on 1989-12-05 with total page 408 pages. Available in PDF, EPUB and Kindle. Book excerpt: At last … a collection of practical protocols for explanting and manipulating neuronal and glial cells. A Dissection and Tissue Culture Manual of the Nervous System Abraham Shahar, Jean de Vellis, Antonia Vernadakis, and Bernard Haber, Editors Among research laboratories involved with neuronal and glial cell cultures and their applications, there is a growing demand for a hand-book describing dissection procedures, culture preparation techniques, and the in vitro manipulation of neural cells and tissues for specific analytical purposes. A Dissection and Tissue Culture Manual of the Nervous System offers a diverse collection of methods that have been developed by and are used routinely within specialized neurobiological laboratories. Written in an easy-to-follow style, the procedures described in this unique guide are designed by experts to be applied by those with limited experience in the field. Organized into ten comprehensive sections, ninety concise contributions from leading laboratories worldwide put forth practical, stepwise protocols for neural cell manipulation and experimentation. Methods encompass: an illustrated outline of techniques for the dissection of brain areas in the fetus and the neonate the dissection of selected specialized structures, such as the ciliary ganglion organotypic. explant culture of nervous tissue dissociated culture of astrocytes, oligodendrocyte, neurons, and Schwann cells reaggregation culture of dissociated cells. Sections devoted to various tissue processing methods and experimental applications of cultured material present histochemical, autoradio-graphic, and immunocytochemical staining and visualization techniques. In situ hybridization methods, as well as preparative procedures for electron microscopy and biochemical and physiological assays, are discussed with an emphasis on methods tailored for the neurobiologist. Alternative techniques for the cultivation of the same organ or cell type from diverse animal species are juxtaposed with a varied selection of methodology and instrumentation, and complemented by key literature citations for further reading, to enable the investigator to chose the appropriate approach for a specific neurobiological application. Presented in a comb-bound format for convenient use on the laboratory bench, A Dissection and Tissue Culture Manual of the Nearvous System will be an essential research companion to graduate students, post-doctoral fellows and other laborabory investigators in cell and developmental neurobiology, neuroanatomy, neurophysiology, neuropharmacology, and biochemistry.

A Dissection and Tissue Culture Manual of the Nervous System

Author :
Publisher :
ISBN 13 : 9780845142820
Total Pages : 371 pages
Book Rating : 4.1/5 (428 download)

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Book Synopsis A Dissection and Tissue Culture Manual of the Nervous System by :

Download or read book A Dissection and Tissue Culture Manual of the Nervous System written by and published by . This book was released on 1989 with total page 371 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Protocols for Neural Cell Culture

Author : Sergey Fedoroff
Publisher : Springer Science & Business Media
ISBN 13 : 1475725868
Total Pages : 284 pages
Book Rating : 4.4/5 (757 download)

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Book Synopsis Protocols for Neural Cell Culture by : Sergey Fedoroff

Download or read book Protocols for Neural Cell Culture written by Sergey Fedoroff and published by Springer Science & Business Media. This book was released on 2013-04-17 with total page 284 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume is designed to be kept close at hand as a ready reference and a guide to laboratory procedures. It is based on tissue culture manuals used for a number of years at international courses on tissue culture at the University of Saskatchewan, made possible by the generous support of the Canadian Council of Animal Care and the Medical Research Council of Canada. Sergey Fedoroff Arleen Richardson The second edition of Protocols for Neural Cell Culture adheres to the prin ciples enunciated in the first edition, but the content has been extensively revised and expanded. Two new chapters have been added to reflect the increased interest in the development and differentiation of the nervous system and in the reconstruction of its circuitry in tissue culture. One chapter deals with slice cul tures in which the organization of the nervous system is preserved. When slice cultures are combined with explant cultures, afferent and efferent projections can be reconstructed. The other chapter deals with aggregating neural cell cul tures, in which "minibrains" can form. Theses are small, uniformly sized spheres of nervous tissue, usually having nerve cells in the center and astrocytes, oligo dendrocytes, and microglia in the periphery. Such cultures can be used to study neutral cell interactions in an organized milieu and for qualitative as well as quantitative studies at biochemical and molecular levels.

The Pharmacology of Nerve and Muscle in Tissue Culture

Author : Alan L. Harvey
Publisher : Springer Science & Business Media
ISBN 13 : 1468488104
Total Pages : 252 pages
Book Rating : 4.4/5 (684 download)

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Book Synopsis The Pharmacology of Nerve and Muscle in Tissue Culture by : Alan L. Harvey

Download or read book The Pharmacology of Nerve and Muscle in Tissue Culture written by Alan L. Harvey and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 252 pages. Available in PDF, EPUB and Kindle. Book excerpt: The techniques of tissue culture were introduced at the beginning of this century. They have become more and more popular as it is realized that they are not as difficult or as esoteric as some early protagonists liked to maintain. Most of the work performed with culture methods has simply concerned cell growth and survival. Biologists have long used culture approaches to provide a simple system in which to study cell division and multiplication. Any pharmacology done on cultured tissue was largely toxicological or as part of a screening programme for poten tial anti-cancer drugs. In the last decade there has been a great increase in the use of excitable cells in tissue culture. Nerves and muscles from a wide variety of sources can maintain their highly differentiated properties in culture. Such cultures offer an attractive preparation for use in physiological and pharmacological investigations. Consequently, a vast amount of work has been produced, and this book is an attempt to review it. It is hoped that this will introduce physiologists and pharmacologists to the potential of culture methods for their experiments and also indicate to more traditional tissue culture users further possible areas of interest. By being more comprehensive in scope and by trying to concentrate largely on drug actions, I hope that the present volume usefully extends the treatment of the subject begun earlier in the excellent works by Crain (1976) and Nelson and Lieberman (1981).

Cell Culture in the Neurosciences

Author : Jane Bottenstein
Publisher : Springer Science & Business Media
ISBN 13 : 1461324734
Total Pages : 392 pages
Book Rating : 4.4/5 (613 download)

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Book Synopsis Cell Culture in the Neurosciences by : Jane Bottenstein

Download or read book Cell Culture in the Neurosciences written by Jane Bottenstein and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 392 pages. Available in PDF, EPUB and Kindle. Book excerpt: A fundamental problem in neuroscience is the elucidation of the cellular and molecular mechanisms underlying the development and function of the nervous system. The complexity of organization, the heteroge neity of cell types and their interactions, and the difficulty of controlling experimental variables in intact organisms make this a formidable task. Because of the ability that it affords to analyze smaller components of the nervous system (even single cells in some cases) and to better control experimental variables, cell culture has become an increasingly valuable tool for neuroscientists. Many aspects of neural development, such as proliferation, differentiation, synaptogenesis, and myelination, occur in culture with time courses remarkably similar to those in vivo. Thus, in vitro methods often provide excellent model systems for investigating neurobiological questions. Ross Harrison described the first culture of neural tissue in 1907 and used morphological methods to analyze the cultures. Since that time the technique has been progressively modified and used to address an ever widening range of developmental questions. In recent years a con vergence of new or improved cell culture, biochemical, electrophysiol ogical, and immunological methods has occurred and been brought to bear on neurobiological questions. This volume is intended not to be comprehensive but rather to highlight some of the latest findings, with a review of previous important work as well, in which combinations of these methods are used.

Neurophysiologic Studies in Tissue Culture

Author : Stanley M. Crain
Publisher :
ISBN 13 :
Total Pages : 300 pages
Book Rating : 4.:/5 (45 download)

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Book Synopsis Neurophysiologic Studies in Tissue Culture by : Stanley M. Crain

Download or read book Neurophysiologic Studies in Tissue Culture written by Stanley M. Crain and published by . This book was released on 1976 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Cell, Tissue, and Organ Cultures in Neurobiology

Author : S. Fedoroff
Publisher : Elsevier
ISBN 13 : 0323143156
Total Pages : 708 pages
Book Rating : 4.3/5 (231 download)

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Book Synopsis Cell, Tissue, and Organ Cultures in Neurobiology by : S. Fedoroff

Download or read book Cell, Tissue, and Organ Cultures in Neurobiology written by S. Fedoroff and published by Elsevier. This book was released on 2012-12-02 with total page 708 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cell, Tissue, and Organ Cultures in Neurobiology emerged from an international workshop held at the University of Saskatchewan in March 1977. This book reviews the uses of cell, tissue, and organ cultures in neurobiological research. It brings together an interdisciplinary perspective from morphology, biochemistry, pharmacology, endocrinology, embryology, and genetics. The book is organized into seven parts. Part I contains papers on the characteristics of differentiated cells. Part II presents studies on cell differentiation in primary cultures. Part III deals with studies on cell cultures and cell strains. Part IV focuses on phenotypic cell expression. Part V examines various cellular interactions. Part VI covers studies on nutrition while Part VII takes up applications of cell tissue and organ cultures in neurobiology. The book is directed toward tissue culturists concerned with the nervous system, as well as all neurobiologists, cell biologists, and embryologists interested in learning how neural cells and tissues behave in cultures and what has been learned about the nervous system using tissue culture methods, including the applicability of tissue cultures to the study of cell differentiation.

Culturing Nerve Cells

Author : Gary Banker
Publisher : MIT Press (MA)
ISBN 13 : 9780262023207
Total Pages : 453 pages
Book Rating : 4.0/5 (232 download)

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Book Synopsis Culturing Nerve Cells by : Gary Banker

Download or read book Culturing Nerve Cells written by Gary Banker and published by MIT Press (MA). This book was released on 1991 with total page 453 pages. Available in PDF, EPUB and Kindle. Book excerpt: This do-it-yourself manual describes complete recipes and protocols for biological scientists, particularly neurobiologists, who want to learn how to grow neurons in culture and how to do it well. Moreover, it provides an understanding of the principles behind the protocols. Offering a more coherent overview of techniques than the usual compendium of methods sections, Culturing Nerve Cells also takes into account the numerous details that can make the difference between success and failure by asking experienced culturists to share their technical expertise. Introductory chapters present a brief course on the nuts and bolts of culturing all types of nerve cells—including the kinds of cultures available and which might be the best for a specific application—methods for cell dissociation, culture media, and substrates, and a description of the techniques needed to characterize and study cultures. More than 20 experts then describe the culture systems that they have developed, including the details of each protocol as well as all of the tricks, lore, and troubleshooting techniques that never appear in the scientific literature. Contents: A User's Guide. Getting Started. General Principles. Types of Nerve Cell Cultures, Their Advantages and Limitations. Primary Dissociated Cell Cultures of Neuronal Tissue. Characterizing and Studying Neuronal Cultures. Culture of Specific Cell Types. Isolated Chick Neurons for the Study of Axonal Growth. Culturing Spinal Neurons and Muscle Cells from Xenopus Embryos. Culturing the Large Neurons of Aplysia. Tissue Culture of Mammalian Autonomic Neurons. Methodologies for the Culture and Experimental Use of the PC12 Rat Pheochromocytoma Cell Line. Cell Culture of Neocortex and Basal Forebrain from Postnatal Rats. Rat Hippocampal Neurons in Low-Density Culture. Cerebellar Cells in Culture. Astroglia in Culture. Tissue Culture Methods for the Study of Myelination. Organotypic Slice Cultures of Neural Tissue.

The Neuron in Tissue Culture

Author : L. W. Haynes
Publisher :
ISBN 13 :
Total Pages : 780 pages
Book Rating : 4.3/5 (91 download)

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Book Synopsis The Neuron in Tissue Culture by : L. W. Haynes

Download or read book The Neuron in Tissue Culture written by L. W. Haynes and published by . This book was released on 1999-02-22 with total page 780 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume in the Handbook Series contains 73 contributions from experts in the range of fields to which neuronal tissue culture may be applied. Twenty-two keynote chapters contain methods, well-illustrated with data from the contributing authors? laboratones, which exemplify ?landmark? applications of the technique. Three further tutorial chapters review artefacts of the tissue culture environment, the understanding and control of neuronal apoptosis in vitro, and the culture substratum. The volume concludes with five chapters containing short protocols describing systematically the maintenance and propagation of neurons from mammalian and avian peripheral and central nervous systems, stem cell populations, invertebrate neurons, and finally selected organised and reconstruction techniques which are given prominent treatment throughout the book. The Neuron in Tissue Culture is a comprehensive reference source dealing exclusively with the tissue culture of neurons. It is essential reading for all those with research interests in neuroscience, including developmental biologists, neurologists, nerve cell physiologists and neuropharmaoologists.

Tissue Culture in Neurobiology

Author : Ezio Giacobini
Publisher :
ISBN 13 :
Total Pages : 544 pages
Book Rating : 4.:/5 (45 download)

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Book Synopsis Tissue Culture in Neurobiology by : Ezio Giacobini

Download or read book Tissue Culture in Neurobiology written by Ezio Giacobini and published by . This book was released on 1980 with total page 544 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Handbook of Innovations in Central Nervous System Regenerative Medicine

Author : António Salgado
Publisher : Elsevier
ISBN 13 : 0128180854
Total Pages : 712 pages
Book Rating : 4.1/5 (281 download)

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Book Synopsis Handbook of Innovations in Central Nervous System Regenerative Medicine by : António Salgado

Download or read book Handbook of Innovations in Central Nervous System Regenerative Medicine written by António Salgado and published by Elsevier. This book was released on 2020-06-10 with total page 712 pages. Available in PDF, EPUB and Kindle. Book excerpt: Handbook of Innovations in CNS Regenerative Medicine provides a comprehensive overview of the CNS regenerative medicine field. The book describes the basic biology and anatomy of the CNS and how injury and disease affect its balance and the limitations of the present therapies used in the clinics. It also introduces recent trends in different fields of CNS regenerative medicine, including cell transplantation, bio and neuro-engineering, molecular/pharmacotherapy therapies and enabling technologies. Finally, the book presents successful cases of translation of basic research to first-in-human trials and the steps needed to follow this path. Areas such as cell transplantation approaches, bio and neuro-engineering, molecular/pharmacotherapy therapies and enabling technologies are key in regenerative medicine are covered in the book, along with regulatory and ethical issues. Describes the basic biology and anatomy of the CNS and how injury and disease affect its balance Discusses the limitations of present therapies used in the clinics Introduces the recent trends in different fields of CNS regenerative medicine, including cell transplantation, bio and neuro-engineering, molecular/pharmacotherapy therapies, and enabling technologies Presents successful cases of translation of basic research to first-in-human trials, along with the steps needed to follow this path

Basic Cell Culture Protocols

Author : Jeffrey W. Pollard
Publisher : Springer Science & Business Media
ISBN 13 : 0896034410
Total Pages : 477 pages
Book Rating : 4.8/5 (96 download)

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Book Synopsis Basic Cell Culture Protocols by : Jeffrey W. Pollard

Download or read book Basic Cell Culture Protocols written by Jeffrey W. Pollard and published by Springer Science & Business Media. This book was released on 1997 with total page 477 pages. Available in PDF, EPUB and Kindle. Book excerpt: Now completely revised and updated from the original, much-acclaimed and bestselling first edition, Basic Cell Culture Protocols, 2nd ed. offers today's most comprehensive collection of easy-to-follow, cutting-edge protocols for the culture of a wide range of animal cells. Its authoritative contributors provide explicit, step-by-step instructions, along with extensive notes and tips that allow both experts and beginners to successfully achieve their desired results. Topics range from basic culture methodology to strategies for culturing previously uncultured cell types and hard-to-culture differentiated cells. Methods are also provided for the analysis of living cells by FACS, video microscopy, and confocal microscopy. Like the first edition, this book should be in every cell culture laboratory and be of use to all who use cell cultures in research.

The Cerebral Circulation

Author : Marilyn J. Cipolla
Publisher : Biota Publishing
ISBN 13 : 1615047239
Total Pages : 82 pages
Book Rating : 4.6/5 (15 download)

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Book Synopsis The Cerebral Circulation by : Marilyn J. Cipolla

Download or read book The Cerebral Circulation written by Marilyn J. Cipolla and published by Biota Publishing. This book was released on 2016-07-28 with total page 82 pages. Available in PDF, EPUB and Kindle. Book excerpt: This e-book will review special features of the cerebral circulation and how they contribute to the physiology of the brain. It describes structural and functional properties of the cerebral circulation that are unique to the brain, an organ with high metabolic demands and the need for tight water and ion homeostasis. Autoregulation is pronounced in the brain, with myogenic, metabolic and neurogenic mechanisms contributing to maintain relatively constant blood flow during both increases and decreases in pressure. In addition, unlike peripheral organs where the majority of vascular resistance resides in small arteries and arterioles, large extracranial and intracranial arteries contribute significantly to vascular resistance in the brain. The prominent role of large arteries in cerebrovascular resistance helps maintain blood flow and protect downstream vessels during changes in perfusion pressure. The cerebral endothelium is also unique in that its barrier properties are in some way more like epithelium than endothelium in the periphery. The cerebral endothelium, known as the blood-brain barrier, has specialized tight junctions that do not allow ions to pass freely and has very low hydraulic conductivity and transcellular transport. This special configuration modifies Starling's forces in the brain microcirculation such that ions retained in the vascular lumen oppose water movement due to hydrostatic pressure. Tight water regulation is necessary in the brain because it has limited capacity for expansion within the skull. Increased intracranial pressure due to vasogenic edema can cause severe neurologic complications and death.

Indwelling Neural Implants

Author : William M. Reichert
Publisher : CRC Press
ISBN 13 : 9781420009309
Total Pages : 296 pages
Book Rating : 4.0/5 (93 download)

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Book Synopsis Indwelling Neural Implants by : William M. Reichert

Download or read book Indwelling Neural Implants written by William M. Reichert and published by CRC Press. This book was released on 2007-12-17 with total page 296 pages. Available in PDF, EPUB and Kindle. Book excerpt: Despite enormous advances made in the development of external effector prosthetics over the last quarter century, significant questions remain, especially those concerning signal degradation that occurs with chronically implanted neuroelectrodes. Offering contributions from pioneering researchers in neuroprosthetics and tissue repair, Indwelling Neural Implants: Strategies for Contending with the In Vivo Environment examines many of these challenges, paying particular attention to how the healing of tissues surrounding an implant can impact the intended use of a device. The contributions are divided into four sections · Part one examines wound healing from the initial insertion trauma through the inflammatory and repair process, explaining how the action of healing varies throughout different areas of the body. · Part two considers various performance issues specific to particular implant components, including those that arise from the chemical, mechanical, thermal, and electrical impact on surrounding tissues. It discusses challenges that result from chronic tissue stimulation and heat effects that occur with on-chip and telemetric processing. · Part three presents both in vitro and in vivo approaches to assessing wound healing response to materials. It includes the contribution of the developer of a chronic hollow fiber membrane implant who explains how an in vivo model is used to assess molecular transport in brain tissue surrounding the implant. · The final section evaluates molecular and materials strategies for intervening in CNS wound repair and enhancing the electrical communication between the electrode surface and the surrounding tissue. It also presents novel approaches to nerve regeneration and repair. This seminal work provides researchers with an up-to-date account of the progress in the field that they can build upon to bring us closer to realizing the full value of neural implants in combating otherwise intractable human health problems.

Central Nervous System Tissue Engineering

Author : A. Wilkinson
Publisher : Springer Nature
ISBN 13 : 3031025822
Total Pages : 112 pages
Book Rating : 4.0/5 (31 download)

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Book Synopsis Central Nervous System Tissue Engineering by : A. Wilkinson

Download or read book Central Nervous System Tissue Engineering written by A. Wilkinson and published by Springer Nature. This book was released on 2022-06-01 with total page 112 pages. Available in PDF, EPUB and Kindle. Book excerpt: Combating neural degeneration from injury or disease is extremely difficult in the brain and spinal cord, i.e. central nervous system (CNS). Unlike the peripheral nerves, CNS neurons are bombarded by physical and chemical restrictions that prevent proper healing and restoration of function. The CNS is vital to bodily function, and loss of any part of it can severely and permanently alter a person's quality of life. Tissue engineering could offer much needed solutions to regenerate or replace damaged CNS tissue. This review will discuss current CNS tissue engineering approaches integrating scaffolds, cells and stimulation techniques. Hydrogels are commonly used CNS tissue engineering scaffolds to stimulate and enhance regeneration, but fiber meshes and other porous structures show specific utility depending on application. CNS relevant cell sources have focused on implantation of exogenous cells or stimulation of endogenous populations. Somatic cells of the CNS are rarely utilized for tissue engineering; however, glial cells of the peripheral nervous system (PNS) may be used to myelinate and protect spinal cord damage. Pluripotent and multipotent stem cells offer alternative cell sources due to continuing advancements in identification and differentiation of these cells. Finally, physical, chemical, and electrical guidance cues are extremely important to neural cells, serving important roles in development and adulthood. These guidance cues are being integrated into tissue engineering approaches. Of particular interest is the inclusion of cues to guide stem cells to differentiate into CNS cell types, as well to guide neuron targeting. This review should provide the reader with a broad understanding of CNS tissue engineering challenges and tactics, with the goal of fostering the future development of biologically inspired designs. Table of Contents: Introduction / Anatomy of the CNS and Progression of Neurological Damage / Biomaterials for Scaffold Preparation / Cell Sources for CNS TE / Stimulation and Guidance / Concluding Remarks