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

ويتم تمويل عمل في المختبر التي تنطوي على زراعة الخلايا العصبية التي Aplysia قائمة المعاهد الوطنية للصحة NIH R01 وR21MH077921.

إعداد (انظر القسم الحلول في نهاية بروتوكول لتكوين حلول) <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إعداد أطباق الثقافة. الزجاج معطف جيدا طبق ماتيك ثقافة أسفل الزجاج مع بولي - L - يسين (صنع في بورات الصوديوم). إضافة كافية تماما لتغطية الزجاج جيدا ويترك ل&gt; 1hr (يمكن ?...

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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>

الإعداد الناجح للثقافات الحسية والحركية وAplysia منحنى التعلم بطيئة نوعا ما ، لأنه ينطوي على تنمية المهارات الحركية الدقيقة المرتبطة microdissection والتلاعب في الخلايا العصبية الفردية ينظر من خلال المجهر ستيريو. في تجربتنا ، فإنه يأخذ 1-3 أسابيع تقريبا من الممارسة للحصو?...

ضرورة إيجاد حلول للثقافة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> 0.35 م 2 MgCl وتخزينها في درجة حرارة الغرفة ، وتستخدم للتخدير. </li><li style=";text-align:right;direction:rtl"> بولي - L - حل ليسين ، سيغما : P - 1524 ، ميغاواط&gt; 300000 ، أدلى في 0.1M البورات الصوديوم pH8.2 0.5mg/ml إلى حل. دوامة كذلك تعقيم وتصفية من خلال 0،22 ميكرون التصفية ، واحفظها في ° 4C. لا تجميد ذوبان الجليد. </li><li style=";text-align:right;direction:rtl"> مسحوق L - 15 متوسطة (يبوفيتش) (سيغما : L4386) تستكمل مع الأملاح على النحو التالي لجعل 1 ليتر <table border="1" cellspacing="0" cellpadding="0" style=";text-align:right;direction:rtl"><tr><td> L - 15 مسحوق </td><td> 13.8g </td></tr><tr><td> كلوريد الصوديوم </td><td> 15.4 ز </td></tr><tr><td> D - غلوكوز </td><td> 6.24 ز </td></tr><tr><td> MgSO 4 7H 2 0 </td><td> 6.45g </td></tr><tr><td> بوكل </td><td> 350 ملغ </td></tr><tr><td> NaHCO 3 </td><td> 170 ملغ </td></tr><tr><td> MgCl 2 6H 2 O </td><td> 5.49 ز </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> إضافة ddH20 إلى 1 لتر. ينبغي أن يكون حول 7،4-7،5 درجة الحموضة ، إضافة 10ML الحل القلم / بكتيريا 100X ، ومن خلال تصفية 0.22 متر تصفية تعقيم. المحل في 4C0 لمدة لا تزيد 1 الشهر. </li><li style=";text-align:right;direction:rtl"> يرصد 1 ٪ البروتياز التاسع (1unit/mg) في L15 (تستكمل مع الأملاح على النحو الوارد أعلاه) أو في ASW فورا قبل الاستخدام ، والتصفية ، من خلال تعقيم ميليبور 0،22 م : حل البروتيني الهضم. وينبغي أن يكون كافيا ل5mls في العقد اثنين من الحيوانات (تأكد مغمورة تماما في حل العقد بروتياز). وذكرت أنها سوف سيجما وقف بيع البروتياز التاسع. A البروتيني بديلا هو : Dispase الثاني (روش كتالوج العلوم التطبيقية # 04942078001). </li><li style=";text-align:right;direction:rtl"> ثقافة المتوسط. مباشرة قبل إعداد الثقافات ، أذاب a قسامة مل 10 من الدملمف ومزجها مع 10 ملل من L15 (تستكمل مع الأملاح على النحو الوارد أعلاه) لجعل 20mls متوسطة الثقافة. إضافة 200 ميكرولتر من 200mm L - الجلوتامين ، واستخدام مزيج جيد لإعداد الثقافات. وينبغي إعداد هذه الوسيلة الجديدة مصنوعة كل الثقافات الوقت. </li><li style=";text-align:right;direction:rtl"> يمكن جعل مياه البحر الاصطناعي من المحيط الفورية (أكواريوم الأنظمة ، معلمه ، OH) ، أو على النحو التالي : 450 مم كلوريد الصوديوم ، و 10 ملي بوكل ، 30 ملم MgCl 2 (6H 2 O) ، و 20 ملي MgSO 4 و 10 ملي CaCl 2 (2H 2 O) ، 10 HEPES مم ، مع تعديل الرقم الهيدروجيني إلى 7.4. </li></ol>

preparation of aplysia sensory-motor neuronal cell cultures

الجهاز العصبي للcalifornica الرخوي البحري Aplysia بسيط نسبيا ، ويتألف من حوالي 20،000 الخلايا العصبية. الخلايا العصبية هي كبيرة (تصل إلى 1 مم في القطر) ومحددة ، ذات أحجام مختلفة ، والأشكال والمواقف وتصبغات ، وتتعرض خارجيا الهيئات خلية في العقد المقترنة five موزعة في جميع أنحاء الجسم من الحيوان. وقد سمحت هذه الخصائص المحققين لتحديد الدوائر الكامنة وراء السلوكيات محددة في هذا الحيوان 1. الاتصال الأحادي المشبك بين الخلايا العصبية الحسية والحركية هي المكون المركزي للمنعكس الخيشومية الانسحاب في الحيوان ، ومنعكس بسيطة الدفاعية التي انسحبت الحيوان الخيشومية في تحفيز استجابة لمسية للسيفون. هذا المنعكس يخضع لأشكال التعلم غير النقابي والجمعياتي ، بما في ذلك التوعية والتعود وتكييف الكلاسيكية. من فائدة خاصة لدراسة اللدونة متشابك ، ويمكن إعادة تشكيل المشبك الحسية والحركية في الثقافة ، حيث تتميز جيدا المحفزات انتزاع أشكال اللدونة التي يرتبط مباشرة في سلوك الحيوان 2،3. على وجه التحديد ، وتطبيق السيروتونين تنتج تعزيز متشابك ذلك ، اعتمادا على بروتوكول التطبيق ، ويستمر لمدة دقيقة (على المدى القصير التيسير) ، وساعات (متوسطة الأجل التيسير) أو أيام (على المدى الطويل التيسير). في المقابل ، تطبيق FMRFamide الارسال الببتيد تنتج ضعف أو الاكتئاب التي متشابك ، اعتمادا على بروتوكول التطبيق ، ويمكن أن تستمر من دقائق إلى أيام (على المدى الطويل والاكتئاب). حجم كبير من الخلايا العصبية يسمح للتسجيل الكهربائي المتكرر للقوة متشابك حادة على مدى فترات من الأيام مع microinjection ناقلات التعبير ، siRNAs ومركبات أخرى لهدف محدد يشير الى شلالات والجزيئات وبالتالي تحديد الخطوات الجزيئية والخلايا البيولوجية التي تكمن وراء التغيرات في فعالية متشابك. ميزة إضافية للنظام الثقافة Aplysia يأتي من حقيقة أن الخلايا العصبية المشبك ، شرح خصوصية 4،5 في الثقافة. وهكذا ، الخلايا العصبية الحسية لا تشكل نقاط الاشتباك العصبي مع أنفسهم (autapses) أو مع غيرها من الخلايا العصبية الحسية ، كما أنها لا تشكل نقاط الاشتباك العصبي مع غير المستهدفة الخلايا العصبية الحركية التي تم تحديدها في الثقافة. ودوالي ، ومواقع للاتصال متشابك بين الخلايا العصبية الحسية والحركية ، وكبيرة بما فيه الكفاية (2-7 ميكرون في القطر) للسماح درس تشكيل المشبك (فضلا عن التغييرات في مورفولوجية متشابك) مع الخلايا العصبية الحركية الهدف على المستوى المجهري الخفيفة. في هذا الفيديو ، ونظهر كل خطوة من إعداد الثقافات الخلايا العصبية الحسية والحركية ، بما في ذلك الكبار والتخدير Aplysia الأحداث ، وتشريح العقد بهم ، والهضم البروتيني من العقد ، وإزالة النسيج الضام التي microdissection ، وتحديد كل من الخلايا العصبية الحسية والحركية ، وإزالة لكل نوع من الخلايا التي microdissection ، والطلاء من الخلايا العصبية الحركية ، بالإضافة إلى ذلك من الخلايا العصبية الحسية والتلاعب neurite الحسية إلى شكل من أشكال الاتصال مع الخلايا العصبية الحركية مثقف.

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

الابتدائي ثقافات<em&gt; Aplysia</em&gt; الحسية والحركية العصبية توفير إعداد نموذج لدراسة تشكيل المشبك واللدونة متشابك<em&gt; في المختبر</em&gt;. هذا الفيديو يوضح تحديد وmicrodissection من الخلايا العصبية الحسية والحركية من<em&gt; Aplysia</em&gt; العقد فضلا عن أساليب لإنشاء وصيانة الخلايا العصبية الحسية والحركية في الثقافة.

إعداد Aplysia الحسية والحركية الثقافات الخلية متعلق بالخلايا العصبية

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

preparation-of-aplysia-sensory-motor-neuronal-cell-cultures

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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 الخلية المستمدة من الثقافات خلية عصبية ظهارية

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 ...

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.

متعلق بالخلايا العصبية الثقافات الأساسي من الأمخاخ في وقت متأخر من المرحلة ذبابة الفاكهة شرانق

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.

التحضير للثقافات متعلق بالخلايا العصبية من أجنة ذبابة الفاكهة المرحلة Midgastrula

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.

التحضير للثقافات ماوس العصبية القشرية نأت

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.

الخلية العصبية من الثقافات Aplysia عن التصوير عالية الدقة من المخاريط النمو

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.

الماوس البشرة الجذعية العصبية خلية كريست (EPI - NCSC) الثقافات

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.

إعداد 2 - dGuo المعاملة الثقافات الجهاز الغدة الصعترية

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.

إعداد وصيانة الخلايا العصبية العقد الجذرية الظهرية في الثقافات Compartmented

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.

التحضير لزراعة الخلايا المكونة للدم الابتدائية من نخاع عظام الفئران لElectroporation

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.

التحضير للثقافات الجرذ الحصى الدماغ العصبية للتنمية والدراسات الدبقية

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