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

Aplysia 뉴런의 culturing 관련된 실험실에서 작업이 목록에 NIH R01와 NIH에 의해 투자 R21MH077921 있습니다.

준비 (솔루션 구성을위한 프로토콜의 끝에있는 솔루션 섹션을 참조) <ol><li> 문화 요리를 준비합니다. 폴리 - L - 라이신 (나트륨 borate에서 만든)로 Mattek 유리 바닥 문화 요리 잘 코트 유리. 완전히 잘 유리를 커버하고&gt; 1 시간 (야간 왼쪽 수)에 남길만큼 추가합니다. 철저하게 폴리 - L - 라이신은 인공 바닷물에서 (ASW) 4-5 번 rinsing하여 제거합니다. 마지막 헹굼 제거한 후, 2 50% L15의 MLS (소금과 보충?...

<ol>
	<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>

그것은 스테레오 현미경을 통해 볼 개별 뉴런의 microdissection 및 조작과 관련된 미세 모터 기술의 개발과 관련된 이후 Aplysia 감각 모터 문화의 성공적인 준비는 다소 느린 학습 곡선이 있습니다. 우리의 경험에서는 모터 뉴런과 감각 뉴런을 쌍으로하는 방법에 대한 자세한 내용은 문화의 건강한 고립 감각 뉴런과 추가 1~3주를 얻는 연습의 약 1-3주 걸립니다. 그 동안 현미경이 culturing (임의의 ...

솔루션은 문화에 필요한 <ol><li> 0.35 M MgCl 2, 실온에서 보관 마취에 사용. </li><li> 폴리 - L - 라이신 솔루션, 시그마 : 0.1M 나트륨 Borate pH8.2로 0.5mg/ml 솔루션에서 만들어진 P - 1524, MW&gt; 300000. 잘하고 필터 소용돌이 필터 0.22 μm의를 통해 소독, 4C의 °에서 그것을 저장합니다. 동결 - 해동하지 마십시오. </li><li> L - 15 중간 분말 (Leibovitz) (시그마 : L4386)는 1리터를 만들기 위해 아래의 염분과 보충 <table border="1" cellspacing="0" cellpadding="0"><tr><td> L - 15 분말 </td><td> 13.8g </td></tr><tr><td> NaCl </td><td> 15.4 g </td></tr><tr><td> D - 포도당 </td><td> 6.24 g </td></tr><tr><td> MgSO 4 7H 2 0 </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 2 시간 2 O </td><td> 1.43g </td></tr><tr><td> HEPES </td><td> 3.53g </td></tr></table> ddH20 1 리터 추가합니다. 산도는 7.4-7.5에 대한 수 100X 펜 / Strep 솔루션 10ml를 추가하고, 0.22 m 필터를 통해 필터 소독해야합니다. 일개월 이상의 이상에 대한 4C0에 보관하십시오. </li><li> 테아제 소화 솔루션 : 1 % 테아제 IX (1unit/mg)는 L15에서 만든 (위의 같은 소금과 보충) 또는 ASW에 즉시 사용하기 전에, 0.22 m Millipore를 통해 필터 살균입니다. 5mls 두 동물 (신경이 완전히 테아제 용액에 잠겨 있는지 확인)에서 신경 충분해야합니다. 시그마들은 프로 테아제 IX 판매를 중단할 것이라고 보도했다. 대체 프로 테아제입니다 Dispase II (로체 응용 과학 카탈로그 # 04942078001). </li><li> 문화 매체. 사전 준비 문화 즉시, hemolymph의 10 ML의 나누어지는를 해동하고 20mls 배지를 만들기 위해 L15 10 MLS (위의 같은 소금과 보충)로 섞는다. L - 글루타민 200mM 200 μl를 추가, 잘 믹스와 문화를 준비하십시오. 때마다 문화가 만들어진 신선한 해당 매체가 준비되어야합니다. </li><li> 인공 해수는 인스턴트 오션 (수족관 시스템, 멘토, OH) 또는 다음과 같이 만들 수 있습니다 : 450 MM NaCl, 10 MM KCl, 30 MM MgCl 2 (6H 2 O), 20 MM MgSO 4, 10 MM CaCl 2 (2 시간 2 O), 7.4로 조정 산도 10 MM HEPES. </li></ol>

preparation of aplysia sensory-motor neuronal cell cultures

해양 연체 동물 Aplysia californica의 신경계는 약 20,000 뉴런으로 구성된, 비교적 간단합니다. 뉴런은 별개의 크기, 형태, 위치 및 pigmentations과 함께, 대형 (직경에 1 ㎜)과 식별되며, 세포 기관은 외부 동물의 체내 분산 오 이점의 신경에 노출됩니다. 이러한 속성은 동물 1 특정 동작을 기본 회로를 윤곽을 그리다하기 위해 조사를 허용합니다. 감각과 운동 뉴런 사이의 monosynaptic 연결 동물의 길 - 철수 반사의 중심 구성 요소, 동물이 사이펀의 촉각 자극에 대한 응답으로의 길을 철회하는 간단한 방어 반사합니다. 이 반사는 sensitization, 요법 이니, 그리고 고전적인 컨디셔닝을 포함하는 비 - 연관과 연관 학습의 형태를 겪습. 잘 특징 자극이 동물 2,3의 행동에 직접적으로 상호를 소성의 형태를 이끌어내는 어디 시냅스 소성의 연구에 특히 이익, 감각 모터 버렸네는 문화의 재구성 수 있습니다. 특히 세로토닌의 응용 프로그램은 시냅스, 응용 프로그램 프로토콜에 따라, (단기 촉진) 분 지속 강화, 시간 (중급 단기 촉진) 또는 일 (장기 촉진) 생산하고 있습니다. 대조적으로, 펩티드 송신기 FMRFamide의 응용 프로그램은 응용 프로그램 프로토콜에 따라 일 분 (장기 우울증)에서 지난 수 시냅스 약화이나 우울증을 생산하고 있습니다. 뉴런의 큰 크기는 표현 벡터, siRNAs 및 특정 신호 폭포와 분자를 대상으로하는 다른 화합물의 microinjection 함께 일 기간 동안 시냅스 강도의 반복 sharp 전극 기록을 허용함으로써 변경 사항을 기초 분자 및 세포 생물 학적 단계를 식별 시냅스 효능 인치 Aplysia 문화 시스템의 추가적인 장점은 뉴런 문화 4,5에서 버렸네 - 특이성을 보여주는 사실에서 비롯됩니다. 따라서, 감각 뉴런은 스스로 (autapses) 또는 다른 감각 뉴런과 시냅스를 형성하지 않으며 그들은 문화가 아닌 대상 확인된 모터 뉴런과 시냅스를 형성 않습니다. varicosities, 감각과 운동 뉴런 사이의 시냅스 접촉 사이트, 대상 모터 뉴런과 버렸네 형성은 (뿐만 아니라 시냅스 형태의 변화) 빛의 미세한 수준에서 공부 할 수 있도록 충분한 (직경 2-7 미크론) 큽니다. 이 비디오에서는, 우리는 그들의 신경, 신경의 테아제 소화, microdissection에 의해 결합 조직의 제거, 감각 및 운동 신경 및 제거 모두의 신분을 해부, anesthetizing 성인 및 청소년 Aplysia 포함한 감각 - 운동 신경 세포의 문화를, 준비의 각 단계를 보여주는 microdissection 각 세포 유형의 모터 신경 세포, 감각 neurite의 감각 신경 세포 및 조작의 이외의 도금은 교양 모터 신경 세포와 접촉을 형성합니다.

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

의 주요 문화<em&gt; Aplysia</em&gt; 감각 - 모터 뉴런은 버렸네 형성하고 시냅스 소성을 연구를위한 모델 준비를 제공합니다<em&gt; 체외에서</em&gt;. 이 비디오에서 감각 및 운동 신경의 식별 및 microdissection을 보여<em&gt; Aplysia</em&gt; 신경뿐만 아니라 문화의 감각 - 모터 뉴런을 설정하고 관리하기위한 방법.

의 준비 Aplysia 감각 - 모터의 연결을 셀 문화

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 세포 - 파생 Neuroepithelial 셀 문화

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 ...

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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.

늦은 단계 Drosophila Pupae의 두뇌에서 기본의 연결을 문화

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.

Midgastrula 스테이지 Drosophila의 태아에서 문화의 연결 준비

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.

Dissociated 마우스 두피 신경 세포 배양의 준비

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.

성장 콘스의 고해상도 이미징을위한 Aplysia에서의 연결을 셀 문화

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.

마우스 표피 신경 크레스트 줄기 세포 (에피 NCSC) 문화

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 - 치료 Thymus 오르간 문화의 준비

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.

Compartmented 문화의 도설 루트 신경의 뉴런의 준비 및 유지 관리

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.

Electroporation에 대한 Murine 골수에서 조혈 세포 차 문화의 준비

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.

신경 세포와 Glia 개발 연구 랫 두뇌 집계 문화의 준비

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