| Spinal Cord Preparation from Adult Red-eared Turtles for Electrophysiological Recordings during Motor Activity
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Author:
Date:
2017-07-05
[Abstract] Although it is known that the generation of movements is performed to a large extent in neuronal circuits located in the spinal cord, the involved mechanisms are still unclear. The turtle as a model system for investigating spinal motor activity has advantages, which far exceeds those of model systems using other animals. The high resistance to anoxia allows for investigation of the fully developed and adult spinal circuitry, as opposed to mammals, which are sensitive to anoxia and where using neonates are often required to remedy the problems. The turtle is mechanically stable and natural ...
[摘要] 虽然已知在位于脊髓的神经元回路中很大程度地进行运动的产生，但是所涉及的机制仍不清楚。乌龟作为调查脊柱运动活动的示范系统具有优势，远远超过使用其他动物的模型系统。对缺氧的高抗性允许对完全发育和成年脊髓电路进行调查，而不是对缺氧敏感的哺乳动物，并且通常需要使用新生儿来补救问题。乌龟是机械稳定的，天然感觉输入可以诱导多种复杂的运动行为，而不需要神经化学物质的应用。在这里，我们提供了如何使成年龟准备的详细方案，也称为电生理调查的综合准备。在这里，通过机械感觉激活，通过细胞内，细胞外和电图记录来记录单细胞和网络活性，可以诱导后肢刮擦反射。该准备工作是由Petersen等人（2014）和Petersen和Berg（2016）以及其他正在进行的研究开发的。
【背景】脊髓电生理学的研究传统上与机械并发症有关，因为许多运动部件和脊柱的灵活性。为了规避这个问题，脊髓经常从柱中解剖出来并移动到可以进行稳定的电生理记录的室。然而，该过程具有缺点，例如，如果要研究多个电动机行为，则适当激活电动机电路是复杂的。此外，缺乏供血和缺乏氧气对电路的健康和完整性有严重影响。一个可以规避所有这些问题的实验模型是龟制剂（Keifer and ...
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| Ex vivo Culture of Adult Mouse Antral Glands
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Author:
Date:
2017-01-05
[Abstract] The tri-dimensional culture, initially described by Sato et al. (2009) in order to isolate and characterize epithelial stem cells of the adult small intestine, has been subsequently adapted to many different organs. One of the first examples was the isolation and culture of antral stem cells by Barker et al. (2010), who efficiently generated organoids that recapitulate the mature pyloric epithelium in vitro. This ex vivo approach is suitable and promising to study gastric function in homeostasis as well as in disease. We have adapted Barker’s protocol to ...
[摘要] 为了分离和表征成年小肠的上皮干细胞，最初由Sato等人（2009）描述的三维培养物已经随后适应于许多不同的器官。其中一个例子是Barker等人（2010）分离和培养窦性干细胞，他们有效地产生了在体外重现成熟幽门上皮的组织细胞。这种“离体”方法是适合的，并且有希望研究体内平衡和疾病中的胃功能。我们已经调整了Barker的方案来比较稳态和再生组织，这里，我们一步一步地仔细地描述了窦腺的分离和培养，以及从细胞分选后可能用于培养的窦腺中分离单细胞一个例子（Fernandez Vallone等人，2016）。
背景来自腺体的小鼠成体干细胞可以在3D matrigel中离体生长，作为“迷你腺体”无限期（Barker等人，2010） 。与EGF，Noggin和R-spondin 1存在下生长的小鼠成年小肠的干细胞相比，胃干细胞需要进一步补充Fgf10，胃泌素，Wnt3a和更高浓度的R-螺旋菌素1（称为作为ENRFGW）获得生产性文化。直到最近，在干细胞消融后，在离体培养系统中成体再生窦腺是否生长，如果是这样，仍然是未知的。使用本方案，证明了内源性和再生的腺体在接种时不会类似地生长并且表现出不同的生长培养要求。
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| Isolation and 3-dimensional Culture of Primary Murine Intestinal Epithelial Cells
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Author:
Date:
2014-05-20
[Abstract] The intestine, together with skin and blood, belongs to the organs with the highest cell turnover, which makes it a perfect model to study cellular processes, such as proliferation and differentiation. Epithelial cell turnover in intestine is possible due to the presence of intestinal stem cells, which are located at the bottom of the crypt. Here, we recapitulate a detailed protocol for the isolation and culture procedures of primary epithelial intestinal cells in a three - dimensional (3D) in vitro system, described for the first time by Hans Clevers group (Sato et al., ...
[摘要] 肠道与皮肤和血液一起属于具有最高细胞更新的器官，这使得它成为研究细胞过程如增殖和分化的完美模型。由于存在位于隐窝底部的肠干细胞，肠上皮细胞更新是可能的。在这里，我们重述了在三维（3D）体外系统中原发性上皮肠细胞的分离和培养程序的详细方案，首次由Hans Clevers组（Sato > et al。，2009）。这种特定的3D培养物保留肠干细胞，其产生分化的子代，例如杯状细胞。该培养物具有许多应用并且代表研究干细胞生物学，上皮细胞再生和移植研究的有用模型。此外，呈现的3D文化可用于调查肠上皮细胞的屏障功能，以及上皮细胞和基质细胞之间的异型细胞相互作用。
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