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从一年生丰年虾胚胎中提取活原肠胚细胞的方案。

Protocol for extracting live blastoderm cells from embryos of annual killifish.

机构信息

Laboratory for Mechanobiology of Transforming Systems, Institute for Biological and Medical Engineering, Schools of Engineering, Medicine and Biological Sciences, Pontificia Universidad Católica de Chile, Santiago 7820436, Chile; Laboratory for Molecular Mechanics of Cell Adhesion, Department of Physiology Pontificia Universidad Católica de Chile, Santiago 8320165, Chile.

Integrative Biology Program, Institute of Biomedical Sciences, Facultad de Medicina, Universidad de Chile, Santiago 8380453, Chile; Biomedical Neuroscience Institute, Santiago 8380453, Chile.

出版信息

STAR Protoc. 2023 Sep 15;4(3):102344. doi: 10.1016/j.xpro.2023.102344. Epub 2023 Jun 22.

DOI:10.1016/j.xpro.2023.102344
PMID:37352104
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10320274/
Abstract

The implementation of in vitro approaches using undifferentiated embryonic cells from annual killifish to complement existing in vivo developmental studies has been hindered by a lack of efficient isolation techniques. Here, we present a protocol to isolate annual killifish blastoderm cells, at the epiboly and early dispersion phase, from embryos. We describe steps for hair removal, embryo cleaning, dechorionation, and cell purification. This protocol may also be used to develop strategies to isolate cells from embryos presenting similar challenges.

摘要

利用一年生食蚊鱼未分化的胚胎细胞进行体外方法的实施，以补充现有的体内发育研究，一直受到缺乏有效分离技术的阻碍。在这里，我们提出了一种从胚胎的胚盘和早期分散阶段分离一年生食蚊鱼胚泡细胞的方案。我们描述了去除毛发、胚胎清洗、去壳和细胞纯化的步骤。该方案也可用于开发从具有类似挑战的胚胎中分离细胞的策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/c6f3102c6095/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/ad4e1ac0571a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/d30ec6510967/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/92f77c442189/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/54fde3acd50d/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/94792a6372fb/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/c6f3102c6095/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/ad4e1ac0571a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/d30ec6510967/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/92f77c442189/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/54fde3acd50d/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/94792a6372fb/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f2c8/10320274/c6f3102c6095/gr5.jpg

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本文引用的文献

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Axis formation in annual killifish: Nodal and β-catenin regulate morphogenesis without Huluwa prepatterning.年度食蚊鱼的体节形成： nodal 和 β-连环蛋白在没有 Huluwa 预图案化的情况下调节形态发生。
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Organization of the Catecholaminergic System in the Short-Lived Fish .短命鱼类中儿茶酚胺能系统的组织
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The nucleus measures shape changes for cellular proprioception to control dynamic cell behavior.核体会测量细胞本体感受的形状变化，以控制细胞的动态行为。
Science. 2020 Oct 16;370(6514). doi: 10.1126/science.aba2644.
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Axis Specification in Zebrafish Is Robust to Cell Mixing and Reveals a Regulation of Pattern Formation by Morphogenesis.斑马鱼中的轴特化对细胞混合具有稳健性，并揭示了形态发生对模式形成的调控。
Curr Biol. 2020 Aug 3;30(15):3063-3064. doi: 10.1016/j.cub.2020.07.022.
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