JLN는 MH 68347에 의해 지원되었다.

hippocampal 분리하기 전에 hippocampal 고립을 시작하기 전에 모든 도구 멸균되어 있는지 확인합니다. 70 % 에탄올과 문화 후드를 스프레이하고, 후드 내부의 도구를 놓으십시오. , 멸균 폴리 - L 코팅 유리 coverslips, pipettors 및 팁, 일회용 pipettes, 그리고 전기 pipettor과 10 cm 배양 접시 - 당신이 6 필요합니다. 이 시점부터는 올바른 살균 기술을 사용해야합니다. <ol><li> 물 목욕 켜고 그것이 37 ° C.까지 가열되어 있는지 확인합니다 </li><li> 4 저장되어있는 다음과 같은 솔루션 ° C가 필요합니다 : <ul><li> 수정된 이글의 중간 (멤) </li><li> Neurobasal </li><li> 행크의 버퍼 생리 식염수 (HBSS) </li><li> Borate 버퍼 솔루션 </li><li> 나트륨 pyruvate 솔루션 </li><li> 증류수에 멸균 필터링 20 % 포도당 용액 </li><li> 후드에서 그들을 배치하기 전에 70 %의 에탄올과 함께 모든 병을을 스프레이하십시오. </li></ul></li><li> 냉동실에서 이러한 냉동 솔루션을 가지고 온수 물 목욕에서 그들을 장소 : <ul><li> 말 혈청 5 ML의 나누어지는 </li><li> B - 27 보충의 한 ML의 나누어지는 </li><li> 100X 항생제의 한 ML의 나누어지는 (페니실린 스트렙토 마이신 플러스) </li><li> L - 글루타민 및 0.5 ML의 나누어지는 </li></ul></li><li> 솔루션은 해동 후, 물 목욕 그들을 데리고 70 % 에탄올과 그들을 스프레이하고, 후드에서 그들을 놓으십시오. 이제 도금과 Neurobasal 매체 준비하실 수 있습니다. </li><li> 해결책이 준비 후 hippocampal 고립을 수행하는 동안 워밍업을 단단히 용기 뚜껑과 37 ° C 목욕에서 그들을 놓으십시오. </li><li> 또한, 냉장고 밖으로 프로 테아제 트립신 (2.5 %)의 나누어지는를 타고 물 욕조에 넣습니다. 트립신은 다음 단계에서 격리됩니다 해부하는 해마를 소화합니다. </li><li> , 15ml 원뿔 튜브를 타고 HBSS로 기입하고, 치료 그룹으로 분류. 그것은 고립된 hippocampi는 다음 단계에서 수집됩니다 여기에 있습니다. </li></ol> Hippocampal 절연 <ol><li> hippocampal 격리를 시작하려면 신생아 새끼는 깨끗해하고 우유 밴드, placentas, 그들의 어머니에 의해 제거 탯줄 코드를 적이 있는지 확인하십시오. </li><li> 청소하고, 후드에서 그들을 곳으로 70 %의 에탄올과 배양 접시에있는 스프레이 새끼를 놓습니다. 동물은 euthanized culturing 바로 앞에 있습니다. </li><li> 자세한 살균 한 접시에서 강아지를 타고 70 % 에탄올로 강아지를 찍어 다음 멸균 HBSS 두 씻는다로. </li><li> 가위로 시체에서 머리를 제거합니다. 같은 가위를 사용하여 피부와 두개골을 통해 했네요. </li><li> 멀리 두뇌에서 두개골 껍질, 괜찮아요 족집게 한 켤레를 사용하여 멸균 HBSS의 작은 금액을 포함하는 작은 페트리 접시에 머리를 놓으십시오. </li><li> 대뇌 반구 다시 필. 해마는 중간 측두엽에있는 작은 해마 모양의 구조입니다. </li><li> 15 ML 튜브의 HBSS 3 ML에 해마과 장소를 제거합니다. 각각의 강아지와 함께 이러한 단계를 반복하고, 15 ML 튜브에 각각 분리된 해마를 놓으십시오. 이제 하나의 세포로 조직을 떼어 놓다 시간이 있습니다. </li></ol> Hippocampal 세포 분리 <ol><li> 모든 hippocampi가 고립되어 후 HBSS와 4.5 ML로 15 ML 튜브를 입력하십시오. </li><li> 후드에있는 물 목욕, 에탄올과 스프레이, 장소에서 트립신을 제거합니다. 튜브에 트립신 0.5 ML를 추가하고 37 15 분 동안 품어 ° C. </li><li> 후드에서 튜브의 바닥에 정착 hippocampi을 방해하지 않도록주의하고, 멸균 피펫으로 튜브에서 HBSS / 트립신 용액을 제거합니다. 부드럽게 관과 소용돌이에 HBSS 5 ML을 추가합니다. ° C 5 분 37 알을 품다. </li><li> 이전 HBSS를 제거하고 신선한 솔루션으로 대체 두번이 단계를 반복합니다. </li><li> 냉동고의 I가 DNAse의 나누어지는 가져가라. 4.5 ML의 HBSS에 hippocampi에 DNase 0.5 ML를 추가합니다. DNAse는 효소 불활 성화를 촉진에 추가됩니다. </li><li> 솔루션을 씹다 (또는 그리고 아래로 피펫)는 동질 때까지. homogenate에 거품을 소개하지 않도록주의하십시오. </li><li> trypan 블루 제외 메서드를 사용하여 세포 생존을 확인합니다. </li><li> 6-8 coverslips을 포함 10cm 페트리 접시를 타고, 도금 매체 10 ML를 추가합니다. coverslips 들어있는 접시에 세포의 원하는 번호를 피펫. 소용돌이는 부드럽게 세포를 분산하고, coverslips가 중복되지 않도록합니다. </li><li> 세포 5 % CO 2 humidified 37 ° C 배양기에서 2-4시간 위해 첨부할 수 있습니다. 세포가 가능한되며 첨부 것을 확인한 후, Neurobasal 매체를 포함하는 각각의 요리에 coverslips을 전송하기만하면됩니다. 인큐베이터에 다시 이런 요리를 놓습니다. </li><li> 일주일에 한 번, 필요한만큼, 신선한 Neurobasal 매체와 실험 치료와 매체의 3 분의 1을 교체하십시오. 이제 문화에서 이러한 세포의 기능 특성 연구로 준비가되어 있습니다. </li></ol> Hippocampal 뉴런의 Fura - 2 칼슘 이미징 <ol><li> 교양 hippocampal 뉴런 48 시간 (하지만 이전)에 대한 체외에 있었 후, 칼슘 이미징 실험을 시작할 수 있습니다. 이 시점에서,이 뉴런들은 프로세스를 확장하기 시작한다. 칼슘 이미징을위한 최적의 연령 범위는 체외 3 일에서 7입니다. </li><li> Fura - 2와 부하 문화 37 30 분 AM ° 그들이 어떤 샘플 매 150ms를 칼슘 표시기 염료 및 512 비트 CCD 카메라를 자극하는 제논 아크 램프를 사용하여 20X 목적에 따라 몇 군데 수있는 후에 C. </li></ol>

<ol>
	<li>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, 2nd Edition. , (1998).</li></ol>

이 프로토콜은 게리 뱅커와 김 고슬린 1의 혁신적인 작품의 수정으로 개발되었습니다. 이 책은 culturing 세포에 관심이 사람을위한 필수적인 리소스입니다 -뿐만 아니라 신경 세포 - 풍부한 문화는 현재 프로토콜 설명했다. 세포 culturing에있는 세 개의 가장 중요한 요소는 다음과 같습니다 불임, 속도, 사용되는 매체의 선택이다. 불임 - 층류 / 살균 후드, 무균 조건 및 무균 인큐베이...

<table><tbody><tr><td>Antibiotic antimitotic</td><td>Invitrogen</td><td>15240062</td><td></td></tr><tr><td>B-27 Supplement</td><td>Invitrogen</td><td>17504044</td><td>Serum free</td></tr><tr><td>Boric Acid</td><td>Sigma-Aldrich</td><td>B6768</td><td></td></tr><tr><td>Dnase I</td><td>Sigma-Aldrich</td><td>DN25</td><td></td></tr><tr><td>Fura-2, AM cell permeant</td><td>Molecular Probes, Life Technologies</td><td>F1221</td><td></td></tr><tr><td>Glucose</td><td>Sigma-Aldrich</td><td>G7528</td><td></td></tr><tr><td>HBSS (10X)</td><td>Invitrogen</td><td>14185052</td><td></td></tr><tr><td>HEPES</td><td>Invitrogen</td><td>15630080</td><td></td></tr><tr><td>L-Glutamine</td><td>Sigma-Aldrich</td><td>G7513</td><td></td></tr><tr><td>MEM</td><td>Invitrogen</td><td>51200038</td><td></td></tr><tr><td>Neurobasal</td><td>Invitrogen</td><td>12348017</td><td>Without phenol red</td></tr><tr><td>Poly-L-Lysine Hydrobromide</td><td>Sigma-Aldrich</td><td>P6282</td><td></td></tr><tr><td>Pyruvic Acid</td><td>Sigma-Aldrich</td><td>P2256</td><td></td></tr><tr><td>Sodium Pyruvate</td><td>Sigma-Aldrich</td><td>P2256</td><td></td></tr><tr><td>Sodium Tetraborate</td><td>Sigma-Aldrich</td><td></td><td></td></tr><tr><td>Trypsin, 2.5% (10X)</td><td>Invitrogen</td><td>15090046</td><td></td></tr></tbody></table>

primary culture of hippocampal neurons from p0 newborn rats

hippocampal 신경의 생리적 특성은 일반적으로 특히 때문에 학습 및 메모리에 해마의 참여의 조사입니다. 기본 hippocampal 세포 culturing은 neuroscientists는 개별 세포와 단일 버렸네 수준에서 뉴런의 활동과 속성을 확인하실 수 있습니다. 이 비디오에서는, 우리는 신생아 쥐의 기본 hippocampal 세포를 분리하고 성장하는 방법을 보여줍니다. 해마는 2~3분만큼 짧은 각 신생아 동물로부터 격리 수 있으며, 문화 두 주에 유지하실 수 있습니다. 우리는 또한 간략하게 ratiometric 칼슘 이미징 이러한 hippocampal 뉴런을 사용하는 방법을 보여줍니다 것입니다. 이 프로토콜은 어떠한 수정을 거의, 해마에 대한 과정을 설명하는 동안, 그것은 두뇌의 다른 지역에 적용할 수 있습니다.

Watch this Scientific Journal Video about Primary Culture of Hippocampal Neurons from P0 Newborn Rats at JoVE.com

Primary Culture of Hippocampal Neurons from P0 Newborn Rats

해부 및 개별 뇌 영역에서 세포의 성장은 세포와 생리적 매개 변수의 조사를 용이하게합니다. 우리는 혈청이없는 환경에서 신경 세포 - 풍부한 문화를 생산하는 주요 세포 culturing에 대한 방법을 설명합니다.

P0 신생아 쥐에서 Hippocampal 신경의 주요 문화

The physiological properties of hippocampal neurons are commonly investigated, especially because of the involvement of the hippocampus in learning and ...

primary-culture-of-hippocampal-neurons-from-p0-newborn-rats

Research

JoVE Journal

Biology

76.5K 조회수.  Michigan State University (MSU). 해부 및 개별 뇌 영역에서 세포의 성장은 세포와 생리적 매개 변수의 조사를 용이하게합니다. 우리는 혈청이없는 환경에서 신경 세포 - 풍부한 문화를 생산하는 주요 세포 culturing에 대한 방법을 설명합니다.

비디오: Primary Culture of Hippocampal Neurons from P0 Newborn Rats

Organotypic Slice Cultures for Studies of Postnatal Neurogenesis

Here we describe a technique for studying hippocampal postnatal neurogenesis in the rodent brain using the organotypic slice culture technique. This method maintains the characteristic topographical morphology of the hippocampus while allowing direct application of pharmacological agents to the developing hippocampal dentate gyrus. Additionally, slice cultures can be maintained for up to 4 weeks and thus, allow one to study the maturation process of newborn granule neurons. Slice cultures allow for efficient pharmacological manipulation of hippocampal slices while excluding complex variables such as uncertainties related to the deep anatomic location of the hippocampus as well as the blood brain barrier. For these reasons, we sought to optimize organotypic slice cultures specifically for postnatal neurogenesis research.

Here we describe a technique for studying hippocampal postnatal neurogenesis using the organotypic slice culture technique. This method allows for in vitro manipulation of adult neurogenesis and allows for the direct application of pharmacological agents to the cultured hippocampus.

출생 후 신경 발생의 연구 Organotypic 슬라이스 문화

Here we describe a technique for studying hippocampal postnatal neurogenesis in the rodent brain using the organotypic slice culture technique. This ...

연구

발생학

Isolation and Culture of Hippocampal Neurons from Prenatal Mice

Primary cultures of rat and murine hippocampal neurons are widely used to reveal cellular mechanisms in neurobiology. By isolating and growing individual neurons, researchers are able to analyze properties related to cellular trafficking, cellular structure and individual protein localization using a variety of biochemical techniques. Results from such experiments are critical for testing theories addressing the neural basis of memory and learning. However, unambiguous results from these forms of experiments are predicated on the ability to grow neuronal cultures with minimum contamination by other brain cell types. In this protocol, we use specific media designed for neuron growth and careful dissection of embryonic hippocampal tissue to optimize growth of healthy neurons while minimizing contaminating cell types (i.e. astrocytes). Embryonic mouse hippocampal tissue can be more difficult to isolate than similar rodent tissue due to the size of the sample for dissection. We show detailed dissection techniques of hippocampus from embryonic day 19 (E19) mouse pups. Once hippocampal tissue is isolated, gentle dissociation of neuronal cells is achieved with a dilute concentration of trypsin and mechanical disruption designed to separate cells from connective tissue while providing minimum damage to individual cells. A detailed description of how to prepare pipettes to be used in the disruption is included. Optimal plating densities are provided for immuno-fluorescence protocols to maximize successful cell culture. The protocol provides a fast (approximately 2 hr) and efficient technique for the culture of neuronal cells from mouse hippocampal tissue.

We provide a protocol for the culture of highly purified hippocampal neurons from prenatal mouse brains without the use of a feeder glial cell layer.

태아 쥐의 Hippocampal 신경 세포의 분리 및 문화

Primary cultures of rat and murine hippocampal neurons are widely used to reveal cellular mechanisms in neurobiology. By isolating and growing individual ...

신경과학

Calcium Phosphate Transfection of Primary Hippocampal Neurons

Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this method on hard-to-transfect cells like primary neurons. Here we describe our detailed protocol for calcium phosphate transfection of hippocampal neurons cocultured with astroglial cells.

차 해마 신경 세포의 칼슘 인산염 형질

Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this ...

Rapid Genotyping of Animals Followed by Establishing Primary Cultures of Brain Neurons

High-resolution analysis of the morphology and function of mammalian neurons often requires the genotyping of individual animals followed by the analysis of primary cultures of neurons. We describe a set of procedures for: labeling newborn mice to be genotyped, rapid genotyping, and establishing low-density cultures of brain neurons from these mice. Individual mice are labeled by tattooing, which allows for long-term identification lasting into adulthood. Genotyping by the described protocol is fast and efficient, and allows for automated extraction of nucleic acid with good reliability. This is useful under circumstances where sufficient time for conventional genotyping is not available, e.g., in mice that suffer from neonatal lethality. Primary neuronal cultures are generated at low density, which enables imaging experiments at high spatial resolution. This culture method requires the preparation of glial feeder layers prior to neuronal plating. The protocol is applied in its entirety to a mouse model of the movement disorder DYT1 dystonia (&#916;E-torsinA knock-in mice), and neuronal cultures are prepared from the hippocampus, cerebral cortex and striatum of these mice. This protocol can be applied to mice with other genetic mutations, as well as to animals of other species. Furthermore, individual components of the protocol can be used for isolated sub-projects. Thus this protocol will have wide applications, not only in neuroscience but also in other fields of biological and medical sciences.

We describe procedures for labeling and genotyping newborn mice and generating primary neuronal cultures from them. The genotyping is rapid, efficient and reliable, and allows for automated nucleic-acid extraction. This is especially useful for neonatally lethal mice and their cultures that require prior completion of genotyping.

동물의 신속한 유전자형은 뇌 신경 세포의 차 문화 구축에 이어

High-resolution analysis of the morphology and function of mammalian neurons often requires the genotyping of individual animals followed by the analysis ...

A Simplified Method for Ultra-Low Density, Long-Term Primary Hippocampal Neuron Culture

Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic hippocampal neurons can often grow successfully on glass coverslips at high density under serum-free conditions, but low density cultures typically require a supply of trophic factors by co-culturing them with a glia feeder layer, preparation of which can be time-consuming and laborious. In addition, the presence of glia may confound interpretation of results and preclude studies on neuron-specific mechanisms. Here, a simplified method is presented for ultra-low density (~2,000 neurons/cm2), long-term (&#62;3 months) primary hippocampal neuron culture that is under serum free conditions and without glia cell support. Low density neurons are grown on poly-D-lysine coated coverslips, and flipped on high density neurons grown in a 24-well plate. Instead of using paraffin dots to create a space between the two neuronal layers, the experimenters can simply etch the plastic bottom of the well, on which the high density neurons reside, to create a microspace conducive to low density neuron growth. The co-culture can be easily maintained for &#62;3 months without significant loss of low density neurons, thus facilitating the morphological and physiological study of these neurons. To illustrate this successful culture condition, data are provided to show profuse synapse formation in low density cells after prolonged culture. This co-culture system also facilitates the survival of sparse individual neurons grown in islands of poly-D-lysine substrates and thus the formation of autaptic connections.

Low density cultures of primary hippocampal neurons usually require glia feeder layer to supply neurotrophic factors and sustain longevity. We describe here a simplified method to culture ultra-low density neurons on glass coverslips in the presence of a high density neuronal feeder layer, which facilitates investigation of specific neuronal-autonomous mechanisms.

매우 낮은 밀도에 대한 단순화 된 방법, 장기 차 해마 신경 세포 문화

Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic ...

Low-Density Primary Hippocampal Neuron Culture

The ability to probe the structure and physiology of individual nerve cells in culture is crucial to the study of neurobiology, and allows for flexibility in genetic and chemical manipulation of individual cells or defined networks. Such ease of manipulation is simpler in the reduced culture system when compared to the intact brain tissue. While many methods for the isolation and growth of these primary neurons exist, each has its own limitations. This protocol describes a method for culturing low-density and high-purity rodent embryonic hippocampal neurons on glass coverslips, which are then suspended over a monolayer of glial cells. This &#39;sandwich culture&#39; allows for exclusive long-term growth of a population of neurons while allowing for trophic support from the underlying glial monolayer. When neurons are of sufficient age or maturity level, the neuron coverslips can be flipped-out of the glial dish and used in imaging or functional assays. Neurons grown by this method typically survive for several weeks and develop extensive arbors, synaptic connections, and network properties.

This article describes the protocol for culturing low-density primary hippocampal neurons growing on glass coverslips inverted over a glial monolayer. The neuron and glial layers are separated by paraffin wax beads. The neurons grown by this method are suitable for high-resolution optical imaging and functional assays.

낮은 밀도 차 해마 신경 세포 문화

The ability to probe the structure and physiology of individual nerve cells in culture is crucial to the study of neurobiology, and allows for flexibility ...

Generation of Neurospheres from Mixed Primary Hippocampal and Cortical Neurons Isolated from E14-E16 Sprague Dawley Rat Embryo

Primary neuron culture is an essential technique in the field of neuroscience. To gain deeper mechanistic insights into the brain, it is essential to have a robust in vitro model that can be exploited for various neurobiology studies. Though primary neuron cultures (i.e., long-term hippocampal cultures) have provided scientists with models, it does not yet represent the complexity of brain network completely. In the wake of these limitations, a new model has emerged using neurospheres, which bears a closer resemblance to the brain tissue. The present protocol describes the plating of high and low densities of mixed cortical and hippocampal neurons isolated from the embryo of embryonic day 14-16 Sprague Dawley rats. This allows for the generation of neurospheres and long-term primary neuron culture as two independent platforms to conduct further studies. This process is extremely simple and cost-effective, as it minimizes several steps and reagents previously deemed essential for neuron culture. This is a robust protocol with minimal requirements that can be performed with achievable results and further used for a diversity of studies related to neuroscience.

Presented here is a protocol for the spontaneous generation of neurospheres enriched in neural progenitor cells from high density plated neurons. During the same experiment, when neurons are plated at a lower density, the protocol also results in prolonged primary rat neuron cultures.

E14-E16 스프라그 Dawley 쥐 배아에서 분리된 혼합 된 기본 해마 및 피질 뉴런에서 신경 구의 생성

Primary neuron culture is an essential technique in the field of neuroscience. To gain deeper mechanistic insights into the brain, it is essential to have ...

Use of Primary Cultured Hippocampal Neurons to Study the Assembly of Axon Initial Segments

Neuronal axon initial segments (AIS) are sites of initiation of action potentials and have been extensively studied for their molecular structure, assembly and activity-dependent plasticity. Giant ankyrin-G, the master organizer of AIS, directly associates with membrane-spanning voltage gated sodium (VSVG) and potassium channels (KCNQ2/3), as well as 186 kDa neurofascin, a L1CAM cell adhesion molecule. Giant ankyrin-G also binds to and recruits cytoplasmic AIS molecules including beta-4-spectrin, and the microtubule-binding proteins, EB1/EB3 and Ndel1. Giant ankyrin-G is sufficient to rescue AIS formation in ankyrin-G deficient neurons. Ankyrin-G also includes a smaller 190 kDa isoform located at dendritic spines instead of the AIS, which is incapable of targeting to the AIS or rescuing the AIS in ankyrin-G-deficient neurons. Here, we described a protocol using cultured hippocampal neurons from ANK3-E22/23-flox mice, which, when transfected with Cre-BFP exhibit loss of all isoform of ankyrin-G and impair the formation of AIS. Combined a modified Banker glia/neuron co-culture system, we developed a method to transfect ankyrin-G null neurons with a 480 kDa ankyrin-G-GFP plasmid, which is sufficient to rescue the formation of AIS. We further employ a quantification method, developed by Salzer and colleagues to deal with variation in AIS distance from the neuronal cell bodies that occurs in hippocampal neuron cultures. This protocol allows quantitative studies of the de novo assembly and dynamic behavior of AIS.

Here, we described a protocol to quantitatively study the assembly and structure of the axon initial segments (AIS) of hippocampal neurons that lack pre-assembled AIS due to the absence of a giant ankyrin-G.

축 사 초기 세그먼트의 조립을 공부 하는 기본 배양 된 해마 뉴런의 사용

Neuronal axon initial segments (AIS) are sites of initiation of action potentials and have been extensively studied for their molecular structure, ...

Easy and Reproducible Low-Density Primary Culture using Frozen Stock of Embryonic Hippocampal Neurons

Neuronal culture is a valuable system for evaluating synaptic functions and drug screenings. In particular, a low-density culture of primary hippocampal neurons allows the study of individual neurons or subcellular components. We have shown subcellular protein localization within a neuron by immunocytochemistry, neuronal polarity, synaptic morphology, and its developmental change using a low-density primary hippocampal culture. Recently, ready-to-use frozen stocks of neurons have become commercially available. These frozen stocks of neurons reduce the time needed to prepare animal experiments and also contribute to the reduction of the number of animals used. Here, we introduce a reproducible low-density primary culture method using a 96-well plate. We used a commercially available frozen stock of neurons from the rat embryonic hippocampus. The neurons can be stably cultured long-term without media changes by reducing the growth of glial cells at particular timepoints. This high-throughput assay using low-density culture allows reproducible imaging-based evaluations of synaptic plasticity.

A ready-to-use frozen stock of neurons is a powerful tool for evaluating synaptic functions. Here, we introduce an easy low-density primary culture from frozen stock using a 96-well plate.

배아 해마 뉴런의 냉동 재고를 사용한 쉽고 재현 가능한 저밀도 1차 배양

Neuronal culture is a valuable system for evaluating synaptic functions and drug screenings. In particular, a low-density culture of primary hippocampal ...

Preparation of Organotypic Slice Cultures from a Rat Pup&#39;s Hippocampus

Source: Mosa, A. J. et al., <a href="https://app.jove.com/v/52353">Organotypic Slice Cultures for Studies of Postnatal Neurogenesis.</a> J. Vis. Exp. (2015).
This video demonstrates a method for preparing organotypic hippocampal slice cultures from the brain of a rat pup. It outlines the steps involved in the dissection of the hippocampus, slicing, and culture conditions required to promote cell viability and growth in the hippocampal slices, ultimately generating organotypic slice cultures.

Preparation of Organotypic Slice Cultures from a Rat Pup's Hippocampus(쥐 새끼의 해마에서 유기형 슬라이스 배양 준비)

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