Name: isolation and culture of dissociated sensory neurons from chick embryos
Uploaded: 2014-09-24T14:00:00.000+00:00
Duration: 11 min 16 s
Description: Watch this Scientific Journal Video about Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos at JoVE.com

نود أن نشكر أليسون Philbrook، بليندا Barbagallo ومايكل فرانسيس للتعليقات الثاقبة على هذه الورقة. وأيد البحث عنها في هذا المنشور من قبل جائزة R15 AREA 1R15NS070172-01A1 منحت لMLL.

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هنا نقدم بروتوكولات مفصلة لعزل وزراعة الخلايا العصبية الحسية فصلها من جنين الفرخ. يولد هذا الإجراء السكان المخصب من الخلايا العصبية المتنامية بقوة في المختبر 7،8،10،16. العديد من التقنيات الخلوية والجزيئية يمكن تطبيقها على هذه الخلايا العصبية مثقف، بما في ?...

الخلايا العصبية هي الخلايا المعقدة التي تحمل المعلومات الأساسية لمجموعة متنوعة من المهام بما في ذلك الإحساس، الرؤية، وحركة السيارات، والتعلم، والذاكرة. فريد من أنواع الخلايا الأخرى، تمتد عمليات الخلايا العصبية تشبه الذراع، ودعا محاور عصبية، لتشكيل العصبية الطرق الأساسية للاتصال. أثناء التطوير المتخصصة المقصورات الموجودة على نصائح من محاور النمو، دعا مخاريط النمو، انتقل من خلال الحفل من العظة خارج الخلية لقيادة محور عصبي إلى وجهتها المناسبة. ليست مفهومة تماما الآليات الجزيئية المعقدة التي تكمن وراء النمو مخروط الملاحة. إلى فهم أفضل لهذه الآليات، استخدمت محققين نماذج زراعة الخلايا لدراسة الخلايا العصبية في بيئة المختبر محددة ومبسطة في. وقد أدى دراسة الخلايا العصبية في الثقافة 1 إلى تقدم كبير في فهمنا لبيولوجيا الخلية العصبية بما في ذلك: تمايز الخلايا العصبية 2، وديناميات هيكل الخلية، الإلتقام والاتجار، التغصناتتنظيم 3،4، تجدد المحاور 5، والحالات السريرية مثل اعتلال الأعصاب 6. بالإضافة إلى ذلك، الخلايا العصبية مثقف قابلة للغاية لمجموعة واسعة من تقنيات البحث بما في ذلك مناعية، خلية السطح المشترك مناعي، لطخة غربية، ترنسفكأيشن، رني، والتصوير الحي مثل تحليل timelapse من مخروط نمو الحركة. وهكذا، زراعة الخلايا العصبية الأولية هو نهج قوي لتوضيح جوانب عديدة من بيولوجيا الخلايا من الخلايا العصبية. ويوفر نموذج خلية ثقافة المحققين مع سيطرة مفصلة على الظروف البيئية والمتغيرات. على سبيل المثال، والطبقات التحتية التي ومطلي الخلايا العصبية (وتنمو عليها) يمكن التلاعب بها بسهولة. هنا، ونحن نقدم تعليمات مفصلة لتوليد اثنين من الطبقات التحتية المتميزة، واحدة مع انخفاض laminin-1 تركيز والآخر مع تشبع تركيزات laminin-1. والمثير للدهشة، أن تركيزات مختلفة من نفس الجزيء لها آثار دراماتيكيةعلى الحالة الداخلية من الخلايا العصبية فضلا عن تكوين سطح الخلية الخاصة بهم. على سبيل المثال، مستويات الخلايا من المخيم ومستويات سطح integrins تختلف اختلافا كبيرا في الخلايا العصبية مطلي على هذين 7،8 الطبقات التحتية. وقد أظهرت دراسات إضافية أن الجزيئات الأخرى، بما في ذلك فبرونيكتين وكبريتات شوندروتن البروتيوغليكان، تأثير التعبير عن جزيئات سطح الخلية والحركة العصبية 7-11. بالإضافة إلى ذلك، جزيئات قابلة للذوبان مثل neurotrophins وneurotropins تؤثر أيضا تكوين غشاء الخلية العصبية والحركة 12-16 ويمكن بسهولة وبدقة التلاعب في نموذج ثقافة الخلية. هنا، نحن تصف طرق لعزل وفصلها الثقافة الخلايا العصبية الحسية من الأجنة الفرخ. وقد استخدم هذا الإجراء لتحقيق اختراقات كبيرة في بيولوجيا الأعصاب، بما في ذلك محور عصبي ثمرة 5،7،8،10،11،16-21 وتم تعديل من إجراء يهدف إلى عزل خلايا العقدة 22. هناكالعديد من المزايا لهذا النهج. أولا، العديد من الميزات من الفرخ عقدة الجذر الظهري (DRG) تطوير تتميز جيدا بما في ذلك الإطار الزمني للولادة، والإرشاد محور عصبي، وملامح تعبير البروتين 2،23-28، وبالتالي توفير أساس المفيد التي تبنى عليها بالمعلومات في تجارب المختبر. الثانية، فصل الثقافات العصبية تسمح المحقق لدراسة الخلايا العصبية أكثر مباشرة مقارنة النهج البديلة باستخدام إإكسبلنتس سليمة DRG (التي تحتوي على الخلايا العصبية والخلايا غير العصبية) و / أو الثقافات المختلطة التي تحتوي على كل من الخلايا العصبية وفصلها الخلايا غير العصبية. ثالثا، الإجراء الموضح هنا واضح ومباشر، وغير مكلفة وقابلة للطلاب الجامعيين. ولذلك، هذه التقنية يمكن استخدامها لأغراض البحث وكذلك لأغراض تعليمية. وعلاوة على ذلك، يجب أن اختلافات طفيفة من هذا البروتوكول يسمح تنقية سريع، عالية الغلة من الخلايا العصبية من مصادر أخرى غير DRGs. على سبيل المثال، يمكن تعديل هذا الإجراء لتوفير neuronally ENRالثقافات iched من الأنسجة الأخرى مثل الدماغ الأمامي الجنينية أو الحبل الشوكي. وقد تم تحسين بروتوكولات مناعية لهذه الثقافات العصبية فصلها ويتم وصفها بالتفصيل هنا. يتم توفير إجراءات مناعية المزدوج ضد جزيء التصاق الخلايا العصبية (NCAM) وintegrins β1. وقد استخدمت البيانات الناتجة من هذه الأساليب immunocytochemical لدراسة الزخرفة المكانية وكثافة عدة الجزيئات في الخلايا العصبية مثقف 8،16.

<table><tbody><tr><td>Sterile small culture dishes (35 mm)</td><td>Corning</td><td>430165</td><td></td></tr><tr><td>Sterile large culture dishes (100 mm)</td><td>Falcon</td><td>353003</td><td></td></tr><tr><td>Sterile large Petri dishes (100 mm)</td><td>VWR</td><td>89000-302</td><td></td></tr><tr><td>Glass Petri dish (100 mm)</td><td>VWR</td><td>D108962</td><td></td></tr><tr><td>Fine forceps</td><td>Fine Science Tools</td><td>11251-10</td><td></td></tr><tr><td>Standard forceps</td><td>Fine Science Tools</td><td>1100-12</td><td></td></tr><tr><td>Coverslips (22 x 22 mm)</td><td>Fisher</td><td>12 518 105K</td><td>0.13-0.17 mm thickness is optimal for microscopy</td></tr><tr><td>Porcelin coverslip holder</td><td>Thomas scientific</td><td>8542E40</td><td></td></tr><tr><td>Fetal Bovine serum</td><td>Gibco</td><td>17502-048</td><td>use 50 ml aliquots per 500 ml of Ham&#039;s F12 media</td></tr><tr><td>HCl</td><td>VWR</td><td>VW3204-1</td><td>use at 2 M, can be used twice before discarding</td></tr><tr><td>NaCl</td><td>Sigma</td><td>S9888</td><td>use 5 M NaCl solution for laminin-1 stock solution</td></tr><tr><td>Laminin-1</td><td>Invitrogen</td><td>23017-015</td><td>Add 72 &amp;micro;l of sterile 5 M NaCl and aliquot into 50 &amp;micro;l</td></tr><tr><td>15 ml Conical tube</td><td>cell treat</td><td>229411</td><td></td></tr><tr><td>PBS CMF 10x</td><td>Gibco</td><td>14200</td><td>used at 1x, dilute with sterile Millipore filtered water</td></tr><tr><td>PBS 10x</td><td>Gibco</td><td>14080</td><td>used at 1x, dilute with sterile Millipore filtered water</td></tr><tr><td>Ham&#039;s F12</td><td>Lonza</td><td>12-615F</td><td></td></tr><tr><td>Fetal Bovine Serum Albumin (BSA)</td><td>Gibco</td><td>10438</td><td>use in F12HS20 at 10%</td></tr><tr><td>HEPES</td><td>Sigma</td><td>H3375</td><td>make sterile 1 M solution in distilled water, dilute in F12 media to obtain final concentration of 10 mM</td></tr><tr><td>Penicillin/streptomyocin</td><td>Sigma</td><td>P0781</td><td>use 5 ml in 500 ml of F12 media</td></tr><tr><td>NT3</td><td>Millipore</td><td>GF031</td><td>Add 1 ml of sterile Millipore water, aliquot under sterile conditions, store at -20 &amp;deg;C, use at final concentration of 10 ng/ml in F12H media, keep in -20 &amp;deg;C non-defrosting freezer</td></tr><tr><td>NGF</td><td>RnD systems</td><td>256-GF</td><td>Add 1 ml of sterile 1x PBS with 0.1% BSA, aliquot under sterile conditions, store at -20 &amp;deg;C, used at final concentration of 10 ng/ml in F12H media</td></tr><tr><td>L-Glutamine</td><td>Sigma</td><td>G7513</td><td>aliquot under sterile conditions, store at -20 &amp;deg;C, use at final concentration of 200 mM in F12H media</td></tr><tr><td>Trypsin</td><td>Sigma</td><td>T4049</td><td>aliquot under sterile conditions, store at -20 &amp;deg;C</td></tr><tr><td>Bovine Serum Albumin (BSA)</td><td>Gibco</td><td>15260</td><td></td></tr><tr><td colspan="4">Other items needed: general dissection instruments, including glass Pasteur pipettes, fertile white leghorn chicken eggs, check egg incubator (humidified, 37 &amp;deg;C), cell incubation chamber (humidified, 37 &amp;deg;C), laminar flow hood, binocular stereovision dissecting scope</td></tr><tr><td colspan="4">&lt;strong&gt;Immunocytochemistry Reagents Table&lt;/strong&gt;</td></tr><tr><td>Sucrose</td><td>Sigma</td><td>S9378</td><td>used in fixative solution (4% paraformaldehye, 30% sucrose, 2x PBS)</td></tr><tr><td>Triton-X 100</td><td>Sigma</td><td>T-8787</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma</td><td>P6148</td><td>used in fixative solution (4% paraformaldehye, 30% sucrose, 2x PBS)</td></tr><tr><td>Fluoromount G</td><td>Southern Biotech</td><td>0100-01</td><td></td></tr><tr><td>Normal goat serum</td><td>Life technologies</td><td>PCN500</td><td></td></tr><tr><td>Microscope slides</td><td>VWR</td><td>16004-430</td><td></td></tr><tr><td colspan="4">&lt;strong&gt;Primary Antibody Table&lt;/strong&gt;</td></tr><tr><td>Antibody against NCAM</td><td>Millipore</td><td>AB5032</td><td>polyclonal</td></tr><tr><td>Antibody against activated &amp;beta;1 ingegrin</td><td>Millipore</td><td>MAB19294</td><td>monoclonal</td></tr><tr><td colspan="4">&lt;strong&gt;Secondary Antibody Table&lt;/strong&gt;</td></tr><tr><td>Goat anti-mouse IgG Alexa 488</td><td>Life technologies</td><td>A11001</td><td></td></tr><tr><td>Goat anti-rabbit IgG Alexa 548</td><td>Life technologies</td><td>A11036</td><td></td></tr></tbody></table>

isolation and culture of dissociated sensory neurons from chick embryos

الخلايا العصبية هي الخلايا متعددة الأوجه التي تحمل المعلومات الأساسية لمجموعة متنوعة من المهام بما في ذلك الإحساس، وحركة السيارات، والتعلم، والذاكرة. يمكن دراسة الخلايا العصبية في الجسم الحي يكون تحديا بسبب تعقيدها، بيئاتها المتنوعة والديناميكية، والقيود الفنية. لهذه الأسباب، ودراسة الخلايا العصبية في المختبر يمكن أن يكون مفيدا لكشف أسرار معقدة من الخلايا العصبية. طبيعة محددة جيدا من نماذج ثقافة الخلية يوفر تحكم مفصلة على الظروف البيئية والمتغيرات. نحن هنا تصف كيفية عزل، فصل، والخلايا العصبية الثقافة الابتدائية من الأجنة الفرخ. هذا الأسلوب هو سريعة وغير مكلفة، ويولد تنمو بقوة الخلايا العصبية الحسية. الإجراء ينتج باستمرار الثقافات التي يتم التخصيب لديها الخلايا العصبية وعدد قليل جدا من الخلايا غير العصبية (أقل من 5٪). الخلايا العصبية الأولية لا تلتزم بشكل جيد للزجاج غير المعالجة أو زراعة الأنسجة البلاستيك، وبالتالي الإجراءات التفصيلية لإنشاء اثنين ديستانط، موصوفة محددة جيدا الطبقات التحتية التي تحتوي على laminin لطلاء الخلايا العصبية. الخلايا العصبية مثقف قابلة للغاية لتقنيات الخلوية والجزيئية متعددة، بما في ذلك التعاون مناعي، تخيل خلية الحية، رني، ومناعية. وقد تم تحسين إجراءات مناعية المزدوج على هذه الخلايا العصبية مثقف وصفها هنا.

بروتوكول الموصوفة هنا تمكن المحققون إلى ثقافة السكان المخصب من فصلها الخلايا العصبية الحسية الجنينية مع عدد قليل جدا من الخلايا (على سبيل المثال، &lt;5٪) غير العصبية 7،8،10،16. ويمكن الحصول على العديد من DRGs القطنية العجزية، الصدر ومناطق عنق الرحم. تبعا للاحتيا?...

Watch this Scientific Journal Video about Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos at JoVE.com

Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos

نماذج ثقافة الخلية توفر تحكم مفصلة على الظروف البيئية وبالتالي توفر منصة قوية لتوضيح جوانب عديدة من بيولوجيا الخلايا العصبية. نحن تصف طريقة سريعة وغير مكلفة، وموثوق بها لعزل وفصل، والثقافة الخلايا العصبية الحسية من الأجنة الفرخ. كما تقدم تفاصيل إعداد الطبقات التحتية ومناعية.

العزلة وثقافة فصل الخلايا العصبية الحسية من الفرخ الأجنة

Neurons are multifaceted cells that carry information essential for a variety of functions including sensation, motor movement, learning, and memory. ...

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Research

JoVE Journal

Neuroscience

14.2K Views.  Assumption College.  نماذج ثقافة الخلية توفر تحكم مفصلة على الظروف البيئية وبالتالي توفر منصة قوية لتوضيح جوانب عديدة من بيولوجيا الخلايا العصبية. نحن تصف طريقة سريعة وغير مكلفة، وموثوق بها لعزل وفصل، والثقافة الخلايا العصبية الحسية من الأجنة الفرخ. كما تقدم ...

Video: Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos

Dissection and Culture of Chick Statoacoustic Ganglion and Spinal Cord Explants in Collagen Gels for Neurite Outgrowth Assays

The sensory organs of the chicken inner ear are innervated by the peripheral processes of statoacoustic ganglion (SAG) neurons. Sensory organ innervation depends on a combination of axon guidance cues1 and survival factors2 located along the trajectory of growing axons and/or within their sensory organ targets. For example, functional interference with a classic axon guidance signaling pathway, semaphorin-neuropilin, generated misrouting of otic axons3. Also, several growth factors expressed in the sensory targets of the inner ear, including Neurotrophin-3 (NT-3) and Brain Derived Neurotrophic Factor (BDNF), have been manipulated in transgenic animals, again leading to misrouting of SAG axons4. These same molecules promote both survival and neurite outgrowth of chick SAG neurons in vitro5,6.
Here, we describe and demonstrate the in vitro method we are currently using to test the responsiveness of chick SAG neurites to soluble proteins, including known morphogens such as the Wnts, as well as growth factors that are important for promoting SAG neurite outgrowth and neuron survival. Using this model system, we hope to draw conclusions about the effects that secreted ligands can exert on SAG neuron survival and neurite outgrowth. 
SAG explants are dissected on embryonic day 4 (E4) and cultured in three-dimensional collagen gels under serum-free conditions for 24 hours. First, neurite responsiveness is tested by culturing explants with protein-supplemented medium. Then, to ask whether point sources of secreted ligands can have directional effects on neurite outgrowth, explants are co-cultured with protein-coated beads and assayed for the ability of the bead to locally promote or inhibit outgrowth. We also include a demonstration of the dissection (modified protocol7) and culture of E6 spinal cord explants. We routinely use spinal cord explants to confirm bioactivity of the proteins and protein-soaked beads, and to verify species cross-reactivity with chick tissue, under the same culture conditions as SAG explants. These in vitro assays are convenient for quickly screening for molecules that exert trophic (survival) or tropic (directional) effects on SAG neurons, especially before performing studies in vivo. Moreover, this method permits the testing of individual molecules under serum-free conditions, with high neuron survival8.

We demonstrate how to dissect and culture chick E4 statoacoustic ganglion and E6 spinal cord explants. Explants are cultured under serum-free conditions in 3D collagen gels for 24 hours. Neurite responsiveness is tested with growth factor-supplemented medium and with protein-coated beads.

تشريح العقدة والثقافة والفرخ Statoacoustic Explants النخاع الشوكي في الهلام الكولاجين لفحوصات ثمرة Neurite

The sensory organs of the chicken inner ear are innervated by the peripheral processes of statoacoustic ganglion (SAG) neurons. Sensory organ innervation ...

Isolation and Culture of Rat Embryonic Neural Cells: A Quick Protocol

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied successfully to the isolation of mouse and human primary neurons and neural progenitors. Dissociated neurons are maintained in serum-free medium up to several weeks. These cultures can be used for nucleofection, immunocytochemistry, nucleic acids preparation, as well as electrophysiology. Older neuronal cultures can also be transfected with a good efficiency rate by lentiviral transduction and, less efficiently, with calcium phosphate or lipid-based methods such as lipofectamine.

We describe a rapid methodology to isolate and culture hippocampal and cortical neurons from rodent embryos. This protocol allows us to perform experiments in which nearly pure neuronal cultures are required.

العزلة والثقافة من خلايا الفئران العصبية الجنينية: بروتوكول سريعة

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied ...

Chicken Embryo Spinal Cord Slice Culture Protocol

Slice cultures can facilitate the manipulation of embryo development both pharmacologically and through gene manipulations. In this reduced system, potential lethal side effects due to systemic drug applications can be overcome. However, culture conditions must ensure that normal development proceeds within the reduced environment of the slice. We have focused on the development of the spinal cord, particularly that of spinal motor neurons. We systematically varied culture conditions of chicken embryo slices from the point at which most spinal motor neurons had been born. We assayed the number and type of motor neurons that survived during the culture period and the position of those motor neurons compared to that in vivo. We found that serum type and neurotrophic factors were required during the culture period and were able to keep motor neurons alive for at least 24 hr and allow those motor neurons to migrate to appropriate positions in the spinal cord. We present these culture conditions and the methodology of preparing the embryo slice cultures using eviscerated chicken embryos embedded in agarose and sliced using a vibratome.

Slice cultures facilitate the manipulation of embryo development by gene and pharmacological perturbations. However, culture conditions must ensure that normal development can proceed within the reduced environment of the slice. We illustrate a protocol that facilitates normal spinal cord development to proceed for at least 24 hr.

الدجاج الأجنة شريحة الحبل الشوكي الثقافة البروتوكول

Slice cultures can facilitate the manipulation of  embryo development both pharmacologically and through gene manipulations. In  this reduced system, ...

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

The marine gastropod mollusk Aplysia californica has a venerable history as a model of nervous system function, with particular significance in studies of learning and memory. The typical preparations for such studies are ones in which the sensory and motoneurons are left intact in a minimally dissected animal, or a technically elaborate neuronal co-culture of individual sensory and motoneurons. Less common is the isolated neuronal preparation in which small clusters of nominally homogeneous neurons are dissociated into single cells in short term culture. Such isolated cells are useful for the biophysical characterization of ion currents using patch clamp techniques, and targeted modulation of these conductances. A protocol for preparing such cultures is described. The protocol takes advantage of the easily identifiable glutamatergic sensory neurons of the pleural and buccal ganglia, and describes their dissociation and minimal maintenance in culture for several days without serum.

Isolation of Sensory Neurons of Aplysia californica for Patch Clamp Recordings of Glutamatergic Currents

We describe the dissection of the nervous system of the marine sea hare Aplysia after anesthesia, the isolation of neurons for short term-tissue culture, and recordings of single cell ion currents via the patch clamp technique.

عزل الخلايا العصبية الحسية من<em&gt; Aplysia californica</em&gt; للحصول على التسجيلات المشبك التصحيح من التيارات الجلوتاميرجي

The marine gastropod mollusk Aplysia californica has a venerable  history as a model of nervous system function, with particular significance in  studies ...

Isolation and Culture of Chick Ciliary Ganglion Neurons

Chick ciliary ganglia (CG) are part of the parasympathetic nervous system and are responsible for the innervation of the muscle tissues present in the eye. This ganglion is constituted by a homogenous population of ciliary and choroidal neurons that innervate striated and smooth muscle fibers, respectively. Each of these neuronal types regulate specific eye structures and functions. Over the years, neuronal cultures of the chick ciliary ganglia were shown to be effective cell models in the study of muscle-nervous system interactions, which communicate through cholinergic synapses. Ciliary ganglion neurons are, in its majority, cholinergic. This cell model has been shown to be useful comparatively to previously used heterogeneous cell models that comprise several neuronal types, besides cholinergic. Anatomically, the ciliary ganglion is localized between the optic nerve (ON) and the choroid fissure (CF). Here, we describe a detailed procedure for the dissection, dissociation and in vitro culture of ciliary ganglia neurons from chick embryos. We provide a step-by-step protocol in order to obtain highly pure and stable cellular cultures of CG neurons, highlighting key steps of the process. These cultures can be maintained in vitro for 15 days and, hereby, we show the normal development of CG cultures. The results also show that these neurons can interact with muscle fibers through neuro-muscular cholinergic synapses.

Chick ciliary ganglia (CG) are part of the parasympathetic nervous system. Neuronal cultures of chick CG neurons were shown to be effective cell models in the study of nerve muscle interactions. We describe a detailed protocol for the dissection, dissociation and in vitro culture of CG neurons from chick embryos.

العزلة والثقافة من الفرخ Ciliary الخلايا العصبية جانجليون

Chick ciliary ganglia (CG) are part of the parasympathetic nervous system and are responsible for the innervation of the muscle tissues present in the ...

Isolation of DRG Neurons and Coculture with Schwann Cell Precursors to Generate Schwann Cells

Source: Tsui, Y, et al. <a href="https://app.jove.com/v/55794">Hypoxic Preconditioning of Marrow-derived Progenitor Cells as a Source for the Generation of Mature Schwann Cells.</a> J. Vis. Exp. (2017).
This video demonstrates a procedure for isolating dorsal root ganglion (DRG) neurons from rat embryos and co-culturing them with Schwann cell precursors to generate mature Schwann cells. The isolated DRGs are processed to isolate neurons and glial cells. The isolated cells are cultured to purify the neurons, which are then co-cultured with Schwann cell-like cells (SCLCs) in a co-culture medium. Interaction with the DRG neurons leads to the maturation of SCLCs into Schwann cells.

عزل الخلايا العصبية DRG والزراعة المشتركة مع سلائف خلايا شوان لتوليد خلايا شوان

Generation of Fate-committed Schwann Cells via Coculture with DRG Neurons
1. Preparation of purified rat DRG neurons

	Autoclave all dissection tools ...

JoVE Encyclopedia of Experiments

Neuronal Culture Techniques

Co-culture and Interaction Studies

Isolation of Cells from Sensory Ganglia

Source: Kaelberer, M. M., et al. <a href="https://app.jove.com/v/53917">A Method to Target and Isolate Airway-innervating Sensory Neurons in Mice.</a> J. Vis. Exp. (2016).
This video demonstrates the isolation of cells from the sensory ganglia of mice. Ganglia are subjected to enzymatic digestion to dissolve the extracellular matrix. Single cells are then collected via filtering through a cell strainer followed by density gradient centrifugation for debris removal. Finally, washing and centrifugation are done to collect the isolated cells for future analysis.

عزل الخلايا عن العقد الحسية

All procedures involving sample collection have been performed in accordance with the institute&#8217;s IRB guidelines.
1. Sensory Ganglia Dissociation

	...

Primary Neuronal Culture

Isolation and Culturing of Cells from Mouse Embryo Dorsolateral Telencephalons

Source: Landeira, B. S., et al. <a href="https://app.jove.com/v/56063">Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System.</a> J. Vis. Exp. (2017)
This video demonstrates a protocol for isolating and culturing primary cerebral cortex cells from the dorsolateral telencephalon of mouse embryos.

عزل واستزراع الخلايا من الدماغ الظهري الجانبي لجنين الفأر

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

Isolating Cranial Neural Folds From a Chick Embryo for a Neural Crest Cell Culture

Source: Jacques-Fricke, B. T., et al. <a href="https://app.jove.com/v/63799">Preparation and Morphological Analysis of Chick Cranial Neural Crest Cell Cultures.</a> J. Vis. Exp. (2022).
This video illustrates a technique for isolating cranial neural folds from the midbrain region of chick embryos. It presents a detailed dissection process of the neural folds, their placement, and attachment on fibronectin-coated coverslips, followed by the observation of neural crest cell migration from the neural folds.

عزل الطيات العصبية القحفية من جنين الفرخ لزراعة خلايا القمة العصبية

Advanced Neuronal Models

Isolating and Culturing Chick Embryonic Neurons on a Multi-Electrode Array

Source: Sanchez, K. R., et al., <a href="https://app.jove.com/v/56300">Assessment of the Effects of Endocrine Disrupting Compounds on the Development of Vertebrate Neural Network Function Using Multi-electrode Arrays.</a> J. Vis. Exp.(2018)
This video demonstrates a method to isolate neurons from the forebrain of an early-stage chick embryo and culture them on a microelectrode array to form a neuronal network.

العزلة وثقافة فصل الخلايا العصبية الحسية من الفرخ الأجنة

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