| In vitro Chondrogenic Hypertrophy Induction of Mesenchymal Stem Cells
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Author:
Date:
2016-12-05
[Abstract] To investigate underlying mechanism of chondrogenic hypertrophy, we need proper in vitro hypertrophic model of mesenchymal stem cells (MSCs). This protocol describes our defined method for induction of in vitro chondrogenic hypertrophy of human umbilical cord blood-derived MSCs (hUCB-MSCs). By adding thyroid hormone (T3; triiodothyronine) and minimum osteogenic-inducing factors to culture medium, we could induce hypertrophy of hUCB-MSCs in vitro. Hypertrophic induction was validated using immunohistochemical analysis, Western blotting and reverse transcriptase polymerase chain reaction.
[摘要] 为了研究软骨形成性肥大的基础机制，我们需要适当的体外间充质干细胞（MSC）的增生模型。该协议描述了我们定义的诱导人脐带血衍生的MSC（hUCB-MSC）的体外软骨形成性肥大的方法。通过将甲状腺激素（T3;三碘甲状腺原氨酸）和最小成骨诱导因子添加到培养基中，我们可以在体外诱导hUCB-MSCs的增生。使用免疫组织化学分析，Western印迹和逆转录酶聚合酶链反应验证肥大性诱导。
关键词：间充质干细胞，体外软骨形成性肥大，成软骨分化，三碘甲腺原氨酸;
[背景] ...
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| Imaging Thick Lymph Node Tissue Sections
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Author:
Date:
2016-09-20
[Abstract] Our protocol describes a simple procedure for imaging thick lymph node sections by 2-photon microscopy. Lymph nodes are sectioned using a vibratome (vibrating microtome) to produce slices of tissue that can then be stained with fluorescently labeled antibodies. The thick tissue sections (150-200 μm depth) allow for the detection of cell clustering that is typically under-represented in thin sections (10-20 μm) used for conventional confocal microscopy. Application of 2-photon microscopy facilitates imaging through the thick volume of the vibratome sections. In combination with automated image processing software, a thick lymph node cross-section image also facilitates quantitation of cellular events within a relatively large area of the tissue, thus providing a clearer picture on the ...
[摘要] 我们的协议描述了一个简单的程序，通过双光子显微镜成像厚淋巴结切片。 使用vibratome（振动切片机）切割淋巴结以产生组织切片，然后可用荧光标记的抗体染色。 厚组织切片（150-200μm深度）允许检测细胞聚集，其通常在用于常规共聚焦显微镜的薄切片（10-20μm）中表现不足。 双光子显微镜的应用有利于成像通过厚的体积的vibratome部分。 与自动图像处理软件组合，厚的淋巴结横截面图像还有助于在组织的相对大的区域内的细胞事件的定量，从而提供关于感兴趣的细胞事件的空间分布的更清晰的图像（例如 。，T细胞聚类）。 该方法也可以容易地应用于其它组织，例如脾或皮肤。
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