Name: generation of bone marrow derived murine dendritic cells for use in 2-photon imaging
Uploaded: 2008-07-09T16:32:00
Description: Watch this Scientific Journal Video about Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging at JoVE.com

美国国立卫生Kirchstein奖学金predoctoral奖学金AI - 64128（MPM）的，GM - 41514（MDC）的GM - 48071（IP）

1）从一个鼠标都股骨骨<ol><li>使用夹层剪刀剪去肌肉和暴露的上方和下方的关节（膝关节和髋关节）的股骨。把握清扫镊子股骨中心和削减的上方和下方的关节离开尽可能完整的骨骺。 </li><li>尽可能使用小夹层剪刀的肌肉清理干净。转移到一盘的RPMI股骨。 </li><li>所有的程序应在油烟机，从这个角度上进行，使用无菌媒体，仪器，移液器和培养皿。 </li></ol> 2）消毒股骨骨头： </p...

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树突状细胞是关键介质的适应性免疫反应和最有效的抗原提呈细胞的特点。文献中对人类和小鼠树突状细胞培养的方法不同类型的细胞因子影响树突状细胞对不同类型的发展。值得注意的是，GM - CSF flt3L，IL4，IL13，TNF -α和IFN -γ是用来在不同的组合，产生成熟的，不成熟的，炎症和稳态像小鼠骨骨髓衍生树突状细胞在体外培养 3-7 。在这里，我们提出了一个简单的方法成熟的小鼠树突状细胞有能力生产归?...

generation of bone marrow derived murine dendritic cells for use in 2-photon imaging

编制小鼠树突状细胞的几种方法可以在文献中发现。在这里，我们提出了一种文化产生大于85％的表面CD11c树突状细胞的方法，回家后皮下注射和目前的抗原，抗原特异性T细胞（见视频）引流淋巴结。此外，我们使用埃森仪器Incucyte跟踪树突状细​​胞的成熟，其中，在第10天，培养细胞的形态是典型的一个成熟的树突状细胞和细胞&lt;85％，是CD11chigh。抗原的外周淋巴结中的演示，通过双光子成像的研究发现，树突状细胞和T细胞相互作用1，2，有三个不同的阶段。第一阶段由高度能动的抗原特异性T细胞和抗原携带树突状细胞的1，2之间的简短的串行接触。第二阶段是标记抗原特异性T细胞和抗原轴承树突状细胞的1，2之间的长期接触。最后，第三阶段的特点是由树突状细胞的分离，恢复活力，并开始划分1，2的T细胞。这是抗原特异性的，可以通过加载抗原细胞跟踪染料标记的树突状细胞的双光子成像分析的交互类型的一个例子。

Watch this Scientific Journal Video about Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging at JoVE.com

Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging

次级淋巴器官树突状细胞的抗原呈现启动T细胞介导的​​适应性免疫反应的关键。在这里，我们展示了文化的骨髓衍生的小鼠树突状细胞，活化，双光子成像标签。

代骨髓衍生的双光子成像使用的小鼠树突状细胞

Several methods for the preparation of murine dendritic cells can be found in the literature. Here, we present a method that produces greater than 85% ...

Research

JoVE Journal

Biology

Dissection and 2-Photon Imaging of Peripheral Lymph Nodes in Mice

夹层和小鼠的外周淋巴结的双光子成像

Two-photon imaging has revealed an elegant choreography of motility and cellular interactions within the lymph node under basal conditions and at the ...

科研

生物学

In situ Imaging of the Mouse Thymus Using 2-Photon Microscopy

在胸腺的鼠标采用双光子显微镜的原位成像

Two-photon Microscopy (TPM) enables us to image deep into the thymus and document the events that are important for thymocyte development.  To follow the ...

A Method for 2-Photon Imaging of Blood Flow in the Neocortex through a Cranial Window

通过颅窗口在新皮层的血流量为双光子成像方法

The ability to image the cerebral vasculature (from large vessels to capillaries) and record blood flow dynamics in the intact brain of living rodents is ...

In Vivo 2-Photon Calcium Imaging in Layer 2/3 of Mice

在体内2光子层钙成像对小鼠的2 / 3

To understand network dynamics of microcircuits in the neocortex, it is essential to simultaneously record the activity of a large number of neurons .  ...

Culture of myeloid dendritic cells from bone marrow precursors

文化髓系树突状细胞从骨髓前体

Myeloid dendritic cells (DCs) are frequently used to study the interactions between innate and adaptive immune mechanisms and the early response to ...

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

主电穿孔的小鼠骨髓造血细胞培养制备

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

Homing of Hematopoietic Cells to the Bone Marrow

归巢的骨髓造血细胞

Homing is the phenomenon whereby transplanted hematopoietic cells are able to travel to and engraft or establish residence in the bone marrow.  Various ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Osteoclast Derivation from Mouse Bone Marrow

小鼠骨髓破骨细胞的推导

Osteoclasts are highly specialized cells that are derived from the monocyte/macrophage lineage of the bone marrow. Their unique ability to resorb both the ...

Expression of Fluorescent Fusion Proteins in Murine Bone Marrow-derived Dendritic Cells and Macrophages

荧光融合蛋白在小鼠骨髓基质树突状细胞和巨噬细胞中的表达

Dendritic cells and macrophages are crucial cells that form the first line of defense against pathogens. They also play important roles in the initiation ...