Name: preparation of aplysia sensory-motor neuronal cell cultures
Uploaded: 2009-06-08T00:00:00
Description: Watch this Scientific Journal Video about Preparation of Aplysia Sensory-motor Neuronal Cell Cultures at JoVE.com

Kültür Aplysia nöron içeren laboratuar İş listesi NIH R01 ve NIH R21MH077921 tarafından finanse edilmektedir.

Hazırlama (bkz. çözümleri bölümünde çözümler kompozisyon için protokol sonunda) <ol><li> Kültür yemekleri hazırlayın. Mattek ile poli-L-lisin (sodyum borat yapılan) cam alt kültür çanak Kat cam. Tamamen cam kapak ve&gt; 1hr (gece sol olabilir) terk etmek yeterli ekleyin. Iyice poli-L-lisin, yapay deniz suyu (ASW) 4-5 kez durulayarak çıkarın. Çıkardıktan sonra, son durulama, 2 ml (tuzları ile desteklenmiş ve 2mm son bir konsantrasyonda L-glutamin içeren)% 50 L15 /% 50 hemolimf ekleyin. Bu k?...

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	<li>Kandel, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+molecular+biology+of+memory+storage:+a+dialogue+between+genes+and+synapses.">The molecular biology of memory storage: a dialogue between genes and synapses.</a> Science. 294 (5544), 1030-1038 (2001).</li><li>Rayport, S. G., Schacher, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+plasticity+in+vitro:+cell+culture+of+identified+Aplysia+neurons+mediating+short-term+habituation+and+sensitization.">Synaptic plasticity in vitro: cell culture of identified Aplysia neurons mediating short-term habituation and sensitization.</a> J Neurosci. 6 (3), 759-763 (1986).</li><li>Goldberg, D. J., Schacher, S., Banker, G., Goslin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Culturing Nerve Cells. , 213-236 (1998).</li><li>Glanzman, D. L., Kandel, E. R., Schacher, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identified+target+motor+neuron+regulates+neurite+outgrowth+and+synapse+formation+of+aplysia+sensory+neurons+in+vitro.">Identified target motor neuron regulates neurite outgrowth and synapse formation of aplysia sensory neurons in vitro.</a> Neuron. 3 (4), 441-450 (1989).</li><li>Lyles, V., Zhao, Y., Martin, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synapse+formation+and+mRNA+localization+in+cultured+Aplysia+neurons.">Synapse formation and mRNA localization in cultured Aplysia neurons.</a> Neuron. 49 (3), 349-356 (2006).</li><li>Martin, K. C., Casadio, A., Zhu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synapse-specific,+long-term+facilitation+of+aplysia+sensory+to+motor+synapses:+a+function+for+local+protein+synthesis+in+memory+storage.">Synapse-specific, long-term facilitation of aplysia sensory to motor synapses: a function for local protein synthesis in memory storage.</a> Cell. 91 (7), 927-938 (1997).</li><li>Guan, Z., Giustetto, M., Lomvardas, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Integration+of+long-term-memory-related+synaptic+plasticity+involves+bidirectional+regulation+of+gene+expression+and+chromatin+structure.">Integration of long-term-memory-related synaptic plasticity involves bidirectional regulation of gene expression and chromatin structure.</a> Cell. 111 (4), 483-493 (2002).</li></ol>

Stereo-mikroskop ile bakıldığında bireysel nöronlar mikrodiseksiyon ve manipülasyon ile ilişkili ince motor becerileri geliştirme içerdiğinden Aplysia duyusal-motor kültürlerin başarılı bir şekilde hazırlanması, biraz daha yavaş bir öğrenme eğrisi vardır. Bizim tecrübelerimize göre, duyusal nöron motor nöron çifti öğrenmek için sağlıklı kültür izole duyu nöronları ve ek olarak 1-3 hafta elde etmek için uygulama yaklaşık 1-3 hafta sürer. Biz rutin olarak bu sürece mikro...

Çözümler kültürü için gerekli <ol><li> 0.35 M MgCl 2, anestezi için kullanılan oda sıcaklığında saklanır. </li><li> Poli-L-lisin çözüm, Sigma: 0.1M Sodyum Borat pH8.2 için 0.5mg/ml çözüm P-1524, MW&gt; 300.000. Bir 0.22 mikron filtre ve filtre Vortex yoluyla sterilize, 4C ° saklayın. Donma-çözülme etmeyin. </li><li> L-15 Orta tozu (Leibovitz) (Sigma: L4386) 1 litre yapmak için aşağıdaki gibi tuzları ile desteklenmiş <table border="1" cellspacing="0" cellpadding="0"><tr><td> L-15 toz </td><td> 13.8g </td></tr><tr><td> NaCl </td><td> 15.4 g </td></tr><tr><td> D-glikoz </td><td> 6,24 g </td></tr><tr><td> MgSO 4 2 0 7H </td><td> 6.45g </td></tr><tr><td> KCl </td><td> 350 mg </td></tr><tr><td> NaHCO 3 </td><td> 170 mg </td></tr><tr><td> MgCl 2 6H 2 O </td><td> 5.49 g </td></tr><tr><td> CaCl 2 2H 2 O </td><td> 1.43g </td></tr><tr><td> Hepes </td><td> 3.53g </td></tr></table> DdH20 için 1 litre ekleyin. PH 7,4-7,5 hakkında 100X Pen / Strep çözüm 10ml ekleyin ve 0,22 m filtre ile filtre sterilize edilmelidir. 1 aydan daha uzun süre saklayın 4C0. </li><li> Proteaz sindirim çözüm:% 1 Proteaz IX (1unit/mg) L15 (yukarıdaki gibi tuzları ile desteklenmiş) ya da ASW kullanımdan hemen önce 0.22 m Millipore aracılığıyla filtre ile sterilize edilmiş. 5mls iki hayvan (ganglia tamamen proteaz çözüm dalmış emin olun) ganglionlar için yeterli olacaktır. Sigma Proteaz IX satan durdurma olacağını bildirdi. Bir yerine proteaz: Dispase II (Roche Uygulamalı Bilimler katalog # 04942078001). </li><li> Kültür ortamı. Hemen hemolimf 10 ml lik bir kısım hazırlanıyor kültürler önce Çözülme ve 20mls kültür ortamı yapmak için L15, 10 ml (yukarıdaki gibi tuzları ile desteklenmiş) ile karıştırın. 200mm L-Glutamin 200 ul ekleyin, iyice karıştırın ve kültürlerin hazırlanması için kullanılacak. Bu ortam, her zaman kültürleri yapılır taze hazırlanmalıdır. </li><li> Yapay Deniz suyu Anında Ocean (Akvaryum Sistemleri, Mentor, OH) ya da şu şekilde yapılabilir: 450 mM NaCl, 10 mM KCl, 30 mM MgCl 2 (6H 2 O), 20 mM MgSO 4, 10 mM CaCl 2 (2H 2 Ç), 7.4 &#39;e ayarlanmış 10 mM pH ile Hepes. </li></ol>

preparation of aplysia sensory-motor neuronal cell cultures

Deniz yumuşakça Aplysia californica sinir sistemi, yaklaşık 20.000 nöronların oluşan, oldukça basittir . Nöronlar, farklı boyutları, şekilleri, konumları ve pigmentations, büyük (çapı 1 mm kadar) ve tanımlanabilir ve hücre organları dışarıdan hayvanın vücut boyunca dağıtılan beş eşleşmiş ganglionlarda maruz kalmaktadır. Bu özellikler araştırmacılar hayvan 1 belirli davranışların altında yatan devreleri tanımlamak için izin verdiler. Duyu ve motor nöronlar arasındaki monosinaptik bağlantı, hayvan solungaç çekilme refleks merkezi bir bileşeni, hayvan sifon dokunsal uyarılara yanıt solungaç çekilmesi basit bir savunma refleksi. Bu refleks hassaslaşması, alışkanlık ve klasik koşullanma da dahil olmak üzere non-ilişkisel ve ilişkisel öğrenme biçimleri, uğrar. Sinaptik plastisite çalışmada özellikle yarar, iyi karakterize uyaranlara 2,3 hayvan davranışlarını doğrudan ilişkilendiren plastisite biçimleri ortaya duyusal-motor sinaps, kültür sulandırılmış olabilir. Özellikle, serotonin uygulama, uygulama protokolü bağlı olarak, (kısa vadeli kolaylaştırma) dakika süren bir sinaptik güçlendirilmesi, saat (orta vadeli kolaylaştırma) veya gün (uzun vadeli kolaylaştırma) üretir. Buna karşılık, peptid verici FMRFamide uygulama uygulama protokolü bağlı olarak, (uzun vadeli depresyonu) gün dakika sürebilir, sinaptik zayıflaması veya depresyon üretir. Büyük boy nöronların sinaptik gücü ifade vektörler, siRNA&#39;lar ve diğer bileşikler, özel sinyalizasyon kaskadlar ve moleküller hedef mikroenjeksiyon ile birlikte gün süre tekrarlanan keskin elektrot kayıt için izin verir ve böylece, değişikliklerin altında yatan moleküler ve hücre biyolojik adımları belirlemek sinaptik etkinliği. Aplysia kültür sistemi ek bir avantajı, nöronlar kültür 4,5 sinaps-özgüllük göstermektedir ki gerçeğinden geliyor . Böylece, duyusal nöronlar edilebilir (autapses) ya da diğer duyusal nöronlar ile sinaps formu değil, ne de kültürü olmayan hedef tespit edilen motor nöronlar ile sinaps formu. Varisler, duyu ve motor nöronlar arasındaki sinaptik temas siteleri, ışık mikroskobik düzeyde incelenmesi gereken hedef motor nöronlar ile sinaps oluşumu (yanı sıra, sinaptik morfoloji değişiklikleri) izin için yeterli (çapı 2-7 mikron) büyük. Bu video, anestezi yetişkin ve çocuk Aplysia dahil olmak üzere, duyusal-motor nöron kültürleri, hazırlama, her adımı gösteren ganglionlar, gangliyon proteaz sindirim, bağ dokusu mikrodiseksiyon kaldırılması, duyu ve motor nöronlar ve kaldırma belirlenmesi diseksiyon mikrodiseksiyon ile her bir hücre tipi, motor nöron, duyusal neurite duyusal nöron ve manipülasyon ek kaplama kültürlü motor nöron iletişim formu.

Watch this Scientific Journal Video about Preparation of Aplysia Sensory-motor Neuronal Cell Cultures at JoVE.com

Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

Primer kültürler<em&gt; Aplysia</em&gt; Duyusal-motor nöronlar sinaps oluşumu ve sinaptik plastisite eğitim için bir model hazırlama<em&gt; In vitro</em&gt;. Bu video duyu ve motor nöronlar tanımlanması ve mikrodiseksiyon gösterir<em&gt; Aplysia</em&gt; Ganglionlar yanı sıra, duyusal-motor nöronların kültür kurmak ve korumak için yöntemler.

Hazırlanması Aplysia Duyusal-motor Nöronal Hücre Kültürleri

The nervous system of the marine mollusk Aplysia californica is relatively simple, consisting of approximately 20,000 neurons.  The neurons are large (up ...

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Preparation of Aplysia Sensory-motor Neuronal Cell Cultures

ES Cell-derived Neuroepithelial Cell Cultures

ES cells have the potential to differentiate into cells from all germ layers, which makes them an attractive tool for the development of new therapies. In general, the differentiation of ES cells follows the concept to first generate immature progenitor cells, which then can be propagated and differentiated into mature cellular phenotypes. This also applies for ES cell-derived neurogenesis, in which the development of neural cells follows two major steps: First, the derivation and expansion of immature neuroepithelial precursors and second, their differentiation into mature neural cells. A common method to produce neural progenitors from ES cells is based on embryoid body (EB) formation, which reveals the differentiation of cells from all germ layers including neuroectoderm. An alternative and more efficient method to induce neuroepithelial cell development uses stromal cell-derived inducing activity (SDIA), which can be achieved by co-culturing ES cells with skull bone marrow-derived stromal cells (1). Both, EB formation and SDIA, reveal the development of rosette-like structures, which are thought to resemble neural tube- and/or neural crest-like progenitors. The neural precursors can be isolated, expanded and further differentiated into specific neurons and glia cells using defined culture conditions. Here, we describe the generation and isolation of such rosettes in co-culture experiments with the stromal cell line MS5 (2-5).

Derivation of neuroepithelial precursors from embryonic stem (ES) cells using stromal cell-derived inducing activity (SDIA).

ES Hücre türetilen nöroepitelyal Hücre Kültürleri

ES cells have the potential to differentiate into cells from all germ layers, which makes them an attractive tool for the development of new therapies. In ...

Biyoloji

Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae

In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. The procedure begins with the removal of brains from animals at 70-78 hrs after puparium formation. The isolated brains are shown after brief incubation in papain followed by several washes in serum-free growth medium. The process of mechanical dissociation of each brain in a 5 ul drop of media on a coverslip is illustrated. The axons and dendrites of the post-mitotic neurons are sheered off near the soma during dissociation but the neurons begin to regenerate processes within a few hours of plating. Images show live cultures at 2 days. Neurons continue to elaborate processes during the first week in culture. Specific neuronal populations can be identified in culture using GAL4 lines to drive tissue specific expression of fluorescent markers such as  GFP or RFP. Whole cell recordings have demonstrated the cultured neurons form functional, spontaneously active cholinergic and GABAergic synapses. A short video segment illustrates calcium dynamics in the cultured neurons using Fura-2 as a calcium indicator dye to monitor spontaneous calcium transients and nicotine evoked calcium responses in a dish of cultured neurons. These pupal brain cultures are a useful model system in which genetic and pharmacological tools can be used to identify intrinsic and extrinsic factors that influence formation and function of central synapses.

This video demonstrates the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. Views of live cultures show neurite outgrowth and imaging of calcium levels using Fura-2.

Geç Stage Drosophila pupa Brain İlköğretim Nöronal Kültürler

In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. The procedure begins with the ...

Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos and their dechorionation using bleach are demonstrated. Using a glass pipet attached to a mouth suction tube, we illustrate the removal of all cells from single embryos. The method for dispersing cells from each embyro into a small (5 l) drop of medium on an uncoated glass coverslip is demonstrated. A view through the microscope at 1 hour after plating illustrates the preferred cell density. Most of the cells that survive when grown in defined medium are neuroblasts that divide one or more times in culture before extending neuritic processes by 12-24 hours. A view through the microscope illustrates the level of neurite outgrowth and branching expected in a healthy culture at 2 days in vitro. The cultures are grown in a simple bicarbonate based defined medium, in a 5% CO2 incubator at 22-24&deg;C. Neuritic processes continue to elaborate over the first week in culture and when they make contact with neurites from neighboring cells they often form functional synaptic connections. Neurons in these cultures express voltage-gated sodium, calcium, and potassium channels and are electrically excitable. This culture system is useful for studying molecular genetic and environmental factors that regulate neuronal differentiation, excitability, and synapse formation/function.

This video demonstrates the preparation of primary neuronal cultures from midgastrula stage Drosophila embryos. Views of live cultures show cells 1 hour after plating and differentiated neurons after 2 days of growth in a bicarbonate-based defined medium. The neurons are electrically excitable and form synaptic connections.

Nöronal Kültürler Midgastrula Sahne Drosophila Embriyolar hazırlanması

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos ...

Preparation of Dissociated Mouse Cortical Neuron Cultures

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures can be used for a variety of applications including immunocytochemistry, biochemistry, electrophysiology, calcium and sodium imaging, protein and/or RNA isolation. These cultures also provide a platform to study the neuronal development of transgenic animals that carry a late embryonic or postnatal lethal gene mutation. The procedure is relatively straight forward, requires some experience in tissue culture technique and should not take longer than two to three hours if you are properly prepared. Careful separation of the cortical rind from the thalamo-cortical fiber tract will reduce the number of unwanted non-neuronal cells. To increase yields of neuronal cells triturate the pieces of the cortical tissue gently after the enzyme incubation step. This is imperative as it prevents unnecessary injury to cells and premature neuronal cell death. Since these cultures are maintained in the absence of glia feeder cells, they also offer an added advantage of growing cultures enriched in neurons.

This video shows a procedure for generating neuronal cultures from late embryo and early postnatal mouse cortex. These cultures can be used for immunocytochemistry, biochemistry, electrophysiology, calcium and sodium imaging and provide a platform to study the neuronal development of transgenic animals that carry a postnatal lethal gene mutation.

Grubuyla Fare Kortikal Nöron Kültürler hazırlanması

This video will guide you through the process for generating cortical neuronal cultures from late embryo and early postnatal mouse brain. These cultures ...

Neuronal Cell Cultures from Aplysia for High-Resolution Imaging of Growth Cones

Neuronal growth cones are the highly motile structures at the tip of axons that can detect guidance cues in the environment and transduce this information into directional movement towards the appropriate target cell. To fully understand how guidance information is transmitted from the cell surface to the underlying dynamic cytoskeletal networks, one needs a model system suitable for live cell imaging of protein dynamics at high temporal and spatial resolution. Typical vertebrate growth cones are too small to quantitatively analyze F-actin and microtubule dynamics. Neurons from the sea hare Aplysia californica are 5-10 times larger than vertebrate neurons, can easily be kept at room temperature and are very robust cells for micromanipulation and biophysical measurements. Their growth cones have very defined cytoplasmic regions and a well-described cytoskeletal system. The neuronal cell bodies can be microinjected with a variety of probes for studying growth cone motility and guidance. In the present protocol we demonstrate a procedure for dissection of the abdominal ganglion, culture of bag cell neurons and setting up an imaging chamber for live cell imaging of growth cones.

Aplysia californica neurons develop large growth cones in culture that are excellent for high-resolution imaging of growth cone motility and guidance. Here, we present a protocol for dissection and plating of Aplysia bag cell neurons as well as for setting up a chamber for live cell imaging.

Büyüme Konileri Yüksek Çözünürlüklü Görüntüleme için Aplysia Nöronal Hücre Kültürleri

Neuronal growth cones are the highly motile structures at the tip of axons that can detect guidance cues in the environment and transduce this information ...

Mouse Epidermal Neural Crest Stem Cell (EPI-NCSC) Cultures

EPI-NCSC are remnants of the embryonic neural crest in an adult location, the bulge of hair follicles. They are multipotent stem cells that have the physiological property to generate a wide array of differentiated cell types, including neurons, nerve supporting cells, smooth muscle cells, bone/cartilage cells and melanocytes. EPI-NCSC are easily accessible in the hairy skin and can be isolated as a highly pure population of stem cells. This video provides a detailed protocol for preparing mouse EPI-NCSC cultures from whisker follicles. The whisker pad of an adult mouse is removed, and whisker follicles dissected. The follicles are then cut longitudinally and subsequently transversely above and below the bulge region. The bulge is removed from the collagen capsule and placed in a culture plate. EPI-NCSC start to emigrate from the bulge explants 3 to 4 days later.

Mouse Epidermal Neural Crest Stem Cell EPI-NCSC Cultures

Here we show our method to isolate mouse epidermal neural crest stem cells (EPI-NCSC). Technique involves micro-dissecting whisker follicles, isolating the bulge and placeing it into tissue culture. EPI-NCSC start to emigrate from bulge explants onto the substratum within 3 - 4 days.

Fare Epidermal Sinir Crest Kök Hücre (EPI-NCSC) Kültürler

EPI-NCSC are remnants of the embryonic neural crest in an adult location, the bulge of hair follicles. They are multipotent stem cells that have the ...

Preparation of 2-dGuo-Treated Thymus Organ Cultures

In the thymus, interactions between developing T-cell precursors and stromal cells that include cortical and medullary epithelial cells are known to play a key role in the development of a functionally competent T-cell pool. However, the complexity of T-cell development in the thymus in vivo can limit analysis of individual cellular components and particular stages of development. In vitro culture systems provide a readily accessible means to study multiple complex cellular processes. Thymus organ culture systems represent a widely used approach to study intrathymic development of T-cells under defined conditions in vitro. Here we describe a system in which mouse embryonic thymus lobes can be depleted of endogenous haemopoeitic elements by prior organ culture in 2-deoxyguanosine, a compound that is selectively toxic to haemopoeitic cells. As well as providing a readily accessible source of thymic stromal cells to investigate the role of thymic microenvironments in the development and selection of T-cells, this technique also underpins further experimental approaches that include the reconstitution of alymphoid thymus lobes in vitro with defined haemopoietic elements, the transplantation of alymphoid thymuses into recipient mice, and the formation of reaggregate thymus organ cultures. (This article is based on work first reported Methods in Molecular Biology 2007, Vol. 380 pages 185-196).

This video demonstrates the dissection and removal of the fetal thymus as well the preparation of ex vivo cultures of 2-dGuo-treated thymus.

2-dGuo Boyalı Thymus Organ Kültürleri hazırlanması

In the thymus, interactions between developing T-cell precursors and stromal cells that include cortical and medullary epithelial cells are known to play ...

Preparation and Maintenance of Dorsal Root Ganglia Neurons in Compartmented Cultures

Neurons extend axonal processes that are far removed from the cell body to innervate target tissues, where target-derived growth factors are required for neuronal survival and function. Neurotrophins are specifically required to maintain the survival and differentiation of innervating sensory neurons but the question of how these target-derived neurotrophins communicate to the cell body of innervating neurons has been an area of active research for over 30 years. The most commonly accepted model of how neurotrophin signals reach the cell body proposes that signaling endosomes carry this signal retrogradely along the axon. In order to study retrograde transport, a culture system was originally devised by Robert Campenot, in which cell bodies are isolated from their axons. The technique of preparing these compartmented chambers for culturing sensory neurons recapitulates the selective stimulation of neuron terminals that occurs in vivo following release of target-derived neurotrophins. Retrograde signaling events that require long-range microtubule dependent retrograde transport have important implications for the treatment of neurodegenerative disorders.

Here we describe the technique of preparing and maintaining compartmented chambers for culturing sensory neurons of the dorsal root ganglia.

Kompartman Kültürleri Dorsal Kök Ganglia Nöronlar hazırlanması ve Bakım

Neurons extend axonal processes that are far removed from the cell body to innervate target tissues, where target-derived growth factors are required for ...

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell  electroporation system and Gene Pulser  electroporation buffer were specifically developed to transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells.This video demonstrates how to establish primary hematopoietic cell cultures from murine bone marrow, and then prepare them for electroporation in the MXcell system. We begin by isolating femur and tibia. Bone marrow from both femur and tibia are then harvested and cultures are established. Cultured bone marrow cells are then transfected and analyzed.

This procedure describes how to establish primary hematopoietic cell cultures from murine bone marrow and is followed by transfection using the Gene Pulser MXCell electroporation system.

Hazırlık Elektroporasyon murin Kemik İliği itibaren İlköğretim Hematopoetik Hücre Kültürleri

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

Preparation of Rat Brain Aggregate Cultures for Neuron and Glia Development Studies

An in vitro system that recapitulates the development and differentiation of progenitors into mature neurons and glia in the central nervous system (CNS) would provide a powerful platform for neuroscientists to investigate axo-glial interactions, properties and differentiation of multipotent progenitors, and progression of oligodendroglial lineage cells at the cellular and molecular level.  We describe here a CNS aggregate culture system from embryonic rat forebrains, which can be maintained in a serum-free medium up to 3-4 weeks and is used in our laboratory as a model to study neuron-glia interaction and CNS myelination.  This video clip will demonstrate how to isolate and grow these CNS aggregate cultures from E16 rat brain.  Furthermore, from the same brain dissection, highly enriched regular dissociated neuronal cultures can be readily obtained and used for various studies on CNS neurons or used for co-cultures with other cells. 

A protocols for an embryonic rat brain aggregate culture system is described. Multipotent progenitors in the aggregates can develop and differentiate into neurons, astrocytes and oligodendrocytes.

Nöron ve Glia Geliştirme Çalışmaları Rat Beyin Agrega Kültürler hazırlanması

An in vitro system that recapitulates the development and differentiation of progenitors into mature neurons and glia in the central nervous system (CNS) ...