Home Technique / Neuroscience / neuron, oligodendrocyte, astrocyte cells culture

Ehlers lab neuron related protocols new recommended protocol
Neuron related protocols from new Ehlers Lab website at Duke University Neuroscience. Protocols include: - Neuron Culture Solutions - Neuron Culturing - Transfecting Neurons with Lipofectamine 2000 - Preparation of Brain Membrane Fractions - Bioti ...

Neural Stem Cell Culture and Cryopreservation new recommended protocol
Open access protocol for neural stem cell culture, its microscopy analysis and cryopreservation from Nature Protocols (Laura Pacey KK et al, McMaster University). The protocol gives detail to isolate and expand neural stem cells in the form of neurospheres fro ...

Hippocampal Neuron Cultures (and mixed cortical cultures) new recommended protocol
A detailed protocol with illustrations on hippocampal neuron culture (by Jean Siao, Ph.D, UNY at Stonybrook). ...

Neuron and astrocyte cell culture protocol new hot protocol
Compiled by Cris Albuquerque (McDermott Lab at Columbia University ). A good guide for preparing Dorsal Horn neuron monocultures, Dorsal Horn and Dorsal Root Ganglion neuron Co-cultures and microisland cultures. (DOC file) ...

Schwann cells culture from adult peripheral nerve protocol
A full accessible paper published at Acta Chururgica Austriaca 30 (Supplement 147): 15-19, 1998 by A.D. Ansselin et al. ...

Primary Culture of Rat Cerebral Astrocytes protocol
PDF thesis on primary culture of rat astrocytes with introduction, methods and materials, discussions and figures. (Kathryn Aleya Weiss) ...

Banker culture of Glial cells protocol
Banker culture of Glial cells Protocol from Meyer lab at Stanford University. ...

Banker Culture Hippocampal Dissection protocol
Banker Culture Hippocampal Dissection Protocol from Meyer lab at Stanford University. (PDF file) ...

Hippocampal co-culture protocol protocol
Meyer lab Hippocampal preparation and co-culture protocol. (PDF file, stanford University). ...

Culture of Dissociated Sympathetic Neurons protocol
A protocol of dissociated sympathetic neurons culture from chicken embryo. (Dr. KA Crutcher, University of Cincinnati) ...

Culture of Dissociated Cortical Neurons protocol
A protocol of dissociated cortical neurons culture from chicken embryo. (Dr. KA Crutcher, University of Cincinnati) ...

Organotypic Hippocampal Slice Cultures protocol
Protocol from Alpha MED Sciences Co., Ltd. Methods of organotypic slice cultures of brain tissue were originally developed in the roller tube (1) and on the semi-permeable membrane (2, 3) and applied sucessfully on the non-permeable multielectrode probe (4). ...

Preparation of a dispersed neuron culture of rat suprachiasmatic nucleus (SCN) protocol
Protocol written by Sato Honma, M.D., Ph.D., Department of Physiology, Hokkaido University Graduate School of Medicine, Japan. (Alpha MED Sciences Co., Ltd.) ...

Protocol for Fluoro-Jade Histofluorescent Labeling of Degenerating Neurons new protocol
This is a detailed protocol on application of fluoro-jade staining on degenerate neurons. (by Dr.Tom Hallam, University of California, Davis) Fluoro-Jade can be used on almost any tissue section. From our experience FJ will stain any tissue section type or ...

Neural Transplant Staining with DiI and Vital Imaging by 2-Photon Laser-Scanning Microscopy new protocol
By Steve M. Potter, Jerome Pine, and Scott E. Fraser, California Institute of Technology. Abstract We are developing a multielectrode silicon "neuroprobe" for maintaining a long-term, specific, two-way electrical interface with nervous tissue. Our approa ...

Rat hippocampal neuron culture protocol from Meyer Lab new protocol
A PDF protocol on rat hippocampal neuron preparation including buffer preparation and general guidlines. (Meyer lab, Standford University) ...

Culture of oligodendrocyte precursor cells (NG2(+)/O1(-)) and oligodendrocytes (NG2(-)/O1(+)) review
Brain Res Brain Res Protoc. 2002 Aug;10(1):23-30. Culture of oligodendrocyte precursor cells (NG2(+)/O1(-)) and oligodendrocytes (NG2(-)/O1(+)) from embryonic rat cerebrum. Itoh K. Department of Molecular Biodynamics, The Tokyo Metropolitan Institute ...

Transfection of primary CNS and PNS neurons by electroporation new review
A pdf original article on neuron electroporation (Cecilia Y. Martinez and Peter J. Hollenbeck, Department of Biological Sciences, Purdue University). Content: I. Introduction II. Electroporation Theory and Principles III. Electroporation Procedure for Ch ...

More neuron protocols

Neuron and glial cell culture protocols
(Curr Protoc Neurosci)
1: Curr Protoc Neurosci. 2006 Aug;Chapter 3:Unit 3.19.

Rodent sensory neuron culture and analysis.

Hall AK.

Case Western Reserve University, Cleveland, Ohio, USA.

Sensory neurons have proven very useful for analysis of neuronal differentiation
in vivo and in vitro. Their utility for in vitro work is based on the fact that
sensory neurons are relatively easy to isolate in large numbers and are amenable
to manipulations in culture. Lumbar ganglia are usually used because their
location in the caudal nervous system means they are the least differentiated at
any developmental stage, allowing the analysis of relatively undifferentiated
cells. Rodent sensory ganglia from embryonic to adult stages can be dissected
effectively and maintained in serum-free medium or in coculture with other cells
or factors. This unit describes generation of embryonic rat lumbar dorsal root
ganglia (DRG) cultures, which form an important model system for investigating
the cellular and molecular mechanisms that regulate neuronal differentiation.
Adult DRG can also be successfully cultured, with a few modifications of the
general protocol.

PMID: 18428634 [PubMed - in process]

2: Curr Protoc Neurosci. 2004 May;Chapter 3:Unit 3.2.

Long-term culture of hippocampal neurons.

Vicario-Abejón C.

Centro de Investigaciones Biológicas, Censejo Superior de Investigaciones
Cientificas, Madrid, Spain.

In culture, hippocampal cells can develop to express neuronal antigens and
acquire mature neuronal morphologies, including axons, complex dendritic trees,
and synapses that are electrophysiologically active. This system is suitable for
studying neuronal differentiation and other events, such as synaptogenesis. It is
also a valuable model for investigating synaptic plasticity and exploring the
mechanisms of neuronal degeneration. This unit provides a protocol for culturing
neurons prepared from embryonic (E-18) rat or mouse hippocampus, but could also
be used to grow neurons from embryonic cortex, olfactory bulb, striatum, or
spinal cord. A second method is included for preparing neuronal cultures from
embryos with different genotypes, such as those from transgenic mice. Also
described is the preparation of polyornithine- and fibronectin-coated coverslips,
which are highly adhesive and promote neurite outgrowth, for use in the culture

PMID: 18428599 [PubMed - in process]

3: Curr Protoc Neurosci. 2001 May;Chapter 6:Unit 6.5.

Acute isolation of neurons from the mature mammalian central nervous system.

Kay AR, Krupa DJ.

University of Iowa, Iowa City, Iowa, USA.

The acute dissociation procedure provides a simple means of isolating neurons
from the mature mammalian central nervous system. The method was primarily
devised to isolate neurons for patch-clamp electrophysiology. It may also prove
useful for single-cell PCR, immunocytochemistry, sorting of fluorescently labeled
cells, or long-term tissue culture of mature neurons. Dissociation is brought
about by a combination of proteolysis and an ionic environment that encourages
breakdown of the tissue. The method allows the isolation of neurons free of glial
ensheathments in as little as 45 min after the sacrifice of the animal. Neurons
so isolated lose fine dendritic branches, although the structure proximal to the
cell body is often maintained, allowing identification of the morphological type
of the neuron. The preparation has the following advantages: (1) the neurons are
fully differentiated; (2) the cells are electronically compact, which improves
the fidelity of the voltage clamp; (3) the cells are removed from the influence
of surrounding cells; and (4) neurons can be isolated from small, circumscribed
loci within the adult central nervous system.

PMID: 18428515 [PubMed - in process]

4: Curr Protoc Neurosci. 2001 May;Chapter 6:Unit 6.11.

Preparation and maintenance of organotypic slice cultures of CNS tissue.

Gähwiler BH, Thompson SM, Muller D.

University of Zurich, Zurich, Switzerland.

Organotypic slice cultures are the in vitro method of choice for applications
requiring long-term survival of the preparation and a high degree of cellular
differentiation and organization resembling that of the original tissue.
Long-term survival is achieved by culturing slices at the air/liquid interface,
either by continuously rotating the preparation (roller-tube cultures) or by
culturing them on semiporous membranes (stationary interface cultures). Both
culture techniques yield nerve cells which are highly differentiated in terms of
their morphological and physiological characteristics. Because most of these
cultures are prepared from 1-week-old postnatal animals, in which the cellular
and tissue organization is already relatively advanced, the original
cytoarchitecture is often remarkably well maintained. Moreover, the presence of a
full complement of glial and nerve cells is thought to provide a microenvironment
facilitating differentiation of neurons. Slice culture also offers unique
advantages for recording from pairs of cells, as a consequence of the high degree
of connectivity between nerve cells. Recently, new applications have emerged such
as the cultivation of slices from knock-out animals with limited postnatal
survival time or alteration of gene expression by viral vectors.

PMID: 18428510 [PubMed - in process]

5: Curr Protoc Neurosci. 2001 May;Chapter 3:Unit 3.7.

Immortalizing central nervous system cells.

Whittemore SR.

University of Miami School of Medicine, Miami, Florida, USA.

This unit presents methods for isolating clonal, neural-derived cell lines. One
approach for isolating such neural cell lines involves a replication-deficient
retrovirus encoding a specific oncogene and a selectable marker which are used to
infect dissociated CNS cells dissected at a developmental stage at which the cell
population of interest has not undergone its terminal mitotic division. Also
presented is a method for cloning by limiting dilution, which may be necessary to
obtain a pure population of cells. Following growth under appropriate selection
conditions, clones are isolated and tested for their ability to differentiate
with the desired phenotypic properties. A method is also provided for coating
tissue culture dishes, which is necessary for successful culture of CNS neurons.

PMID: 18428471 [PubMed - in process]

6: Curr Protoc Neurosci. 2001 May;Chapter 3:Unit 3.3.

Culture of substantia nigra neurons.

Studer L.

National Institute of Neurological Disorders and Stroke, Bethesda, Maryland, USA.

Primary cultures of nigral tissue are widely used as a model system to assay
effects of trophic and toxic agents on dopaminergic neurons. Cultured
dopaminergic neurons have been successfully transplanted in animals and led to
behavioral improvement in animal models of Parkinson's disease. Cell cultures
have also been used to study the development of substantia nigra, allowing
investigators to identify early inductive events important for nigral development
and to study dopaminergic differentiation and target innervation. This unit
provides simple and reliable culture protocols for these applications. The first
approach presented is the preparation of dissociated nigral cell cultures, the
later steps of which can be used as a simple and efficient assay for testing
growth factors. A second approach is the preparation of free-floating roller tube
cultures, which may be used as a tool for neural transplantation and to study
more complex developmental events. A third approach is the production of
organotypic cultures using chicken plasma as a matrix. Organotypic cultures can
maintain the in vivo cytoarchitecture of a host region in vitro.

PMID: 18428467 [PubMed - in process]

How to grow primary neurons
(Neuroscience forum)
astrocyte-neuron co-culture

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