Issue Information

Q3 Biochemistry, Genetics and Molecular Biology Current Protocols in Cell Biology Pub Date : 2020-09-04 DOI:10.1002/cpcb.92
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Abstract

Cover: In Duband et al. (http://doi.org/10.1002/cpcb.109), the image shows Phenotypic characterization of neural crest cell cultures after several days in culture with cell lineage markers. (A) Phase contrast overview of a 48-hr neural tube explant, showing, on the left, the dense network of neurites (arrows) emerging out of the ventral side of the neural tube and, on the right, the presence of cells with distinct morphologies and shapes among the neural crest cell population. (B) Detailed views of the main cell types derived from the neural crest outgrowth after 3 or 5 days in culture: Panel 1, large, well spread myofibroblasts; Panel 2, neurons with long neurites sitting on top of neural crest cells; and Panel 3, little, aster-shaped melanoblasts. Note that these cells are often assembled as separate colonies. (C) Immunofluorescence labeling of neural crest-derived cell populations after 3 days in culture with antibodies to Sox-10, HNK-1, α-SMA, βIII-tubulin (Tuj-1), and Mitf revealed by secondary antibodies conjugated to Alexa-488 (green), Cy-3 (red), and Cy-5 (purple in C1-C3 and yellow in C4). Nuclei are visualized with Hoechst staining (blue). Panel 1 shows a group of myofibroblasts characterized by a dense meshwork of α-SMA in their cytoplasm. Most of them do not express Sox-10 in their nuclei and HNK-1 on their surface but a few cells corresponding possibly to myofibroblasts having not yet completed their differentiation process express all three markers together (arrows). Besides myofibroblasts, a few large cells with an astrocyte-like shape and expressing both HNK1 and Sox-10 represent early differentiating glial cells (arrowheads). Panel 2 shows neurons expressing βIII-tubulin but no Sox-10. Panel 3 shows a large group of melanoblasts expressing MITF in their nuclei but no HNK-1. As for myofibroblasts, a few melanoblasts can be seen coexpressing Mitf and HNK-1 (arrows), likely corresponding to early melanoblasts. Panel 4 shows persistent Sox-10 positive, HNK-1 positive undifferentiated neural crest cell progenitors in the outgrowth. Bars in A, B, and C = 100 µm.

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封面:在Duband等人(http://doi.org/10.1002/cpcb.109)中,图像显示了在细胞谱系标记培养几天后神经嵴细胞培养的表型特征。(A) 48小时神经管外植体的相衬图,左侧显示神经管腹侧出现密集的神经突网络(箭头),右侧显示神经嵴细胞群中存在形态和形状不同的细胞。(B)培养3或5天后神经嵴生长的主要细胞类型的详细视图:第1组,大的,扩散良好的肌成纤维细胞;图2,长突位于神经嵴细胞顶部的神经元;第三组,小的,星形的黑色素母细胞。注意,这些细胞通常作为独立的菌落聚集在一起。(C)培养3天后神经嵴源性细胞群的免疫荧光标记,其中有针对ox-10、HNK-1、α-SMA、β iii -微管蛋白(Tuj-1)和Mitf的抗体,这些抗体由结合了Alexa-488(绿色)、Cy-3(红色)和Cy-5 (C1-C3为紫色,C4为黄色)的二抗显示。赫斯特染色显示细胞核(蓝色)。图1显示一组肌成纤维细胞,其细胞质中有致密的α-SMA网状结构。它们中的大多数在细胞核中不表达Sox-10,在表面不表达HNK-1,但少数可能与肌成纤维细胞相对应的尚未完成分化过程的细胞同时表达这三种标记(箭头)。除肌成纤维细胞外,少数具有星形细胞样形状且同时表达HNK1和Sox-10的大细胞代表早期分化的胶质细胞(箭头)。图2显示神经元表达β iii -微管蛋白,但不表达Sox-10。图3显示一大群黑素母细胞在细胞核中表达MITF,但不表达HNK-1。对于肌成纤维细胞,可以看到少数黑色素母细胞共表达Mitf和HNK-1(箭头),可能与早期的黑色素母细胞相对应。图4显示出生长物中持续存在Sox-10阳性、HNK-1阳性的未分化神经嵴细胞祖细胞。A、B、C中的棒材= 100µm。
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Current Protocols in Cell Biology
Current Protocols in Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology
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期刊介绍: Developed by leading scientists in the field, Current Protocols in Cell Biology is an essential reference for researchers who study the relationship between specific molecules and genes and their location, function and structure at the cellular level. Updated every three months in all formats, CPCB is constantly evolving to keep pace with the very latest discoveries and developments.
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