C.エレガンス 芽球体解剖:卵殻を除去し、胚細胞を解離する方法

Published: April 30, 2023

Abstract

出典:スー、C.R.、他.孤立したカエノハブディティスエレガンス胚芽芽胞子と粘着ポリスチレンビーズを用いた空間細胞接触パターンの体外再構成。J. ヴィス・エクス(2019). 

このビデオでは、初期胚から C.エレガンス ブラストミアを単離する方法を紹介します。 得られた細胞は、細胞培養または ex vivo 実験に適している。

Protocol

このプロトコルは、   単離されたカエノハブディティスエレガンス胚芽胚芽球と粘着ポリスチレンビーズ、J.Vis.Exp.(2019)を用いた空間細胞接触パターンのインビトロ再構成からの抜粋である。 胚芽胞体の単離 漂白液に触れないように手袋とラ?…

Materials

Aspirator Tube Assembly Drummond 21-180-13 For the blastomere isolation.
Caenorhabditis elegans strain: N2, wild-type Caenorhabditis Genetics Center N2 Strain used in this study
Caenorhabditis elegans strain: KSG5, genotype: zuIs45; itIs37 in house KSG5 Strain used in this study
Calibrated Mircopipets, 10 µL Drummond 21-180-13 For the blastomere isolation
CD Lipid Concentrate Life Technologies 11905031 For the blastomere isolation. Work in the tissue culture hood.
Clorox Clorox N. A. For the blastomere isolation. Open a new bottle when the hypochlorite treatment does not work well.
Coverslip holder In house N.A. For the blastomere isolation.
Dissecting microscope: Zeiss Stemi 508 with M stand. Source of light is built-in LED. Magnification of eye piece is 10X. Carl Zeiss Stemi 508 For the blastomere isolation.
Fetal Bovine Serum, Qualified One Shot, Canada origin Gibco A3160701 For the blastomere isolation. Work in the tissue culture hood.
General Use and Precision Glide Hypodermic Needles, 25 gauge BD 14-826AA For the blastomere isolation
Inulin Alfa Aesar AAA1842509 For the blastomere isolation
MEM Vitamin Solution (100x) Gibco 11120052 For the blastomere isolation.
Multitest Slide 10 Well MP Biomedicals ICN6041805 For the blastomere isolation
Penicillin-Streptomycin (10,000 U/mL) Gibco 15140148 For the blastomere isolation.
Polyvinylpyrrolidone Fisher BioReagents BP431-100 For the blastomere isolation
Potassium Chloride Bioshop POC888 For the blastomere isolation
Schneider’s Drosophila Sterile Medium Gibco 21720024 For the blastomere isolation. Work in the tissue culture hood.
Sodium Chloride Bioshop SOD001 For the blastomere isolation
Sodium Hydroxide Solution, 10 N Fisher Chemical SS255-1 For the blastomere isolation
Syringe Filters, PTFE, Non-Sterile Basix 13100115 For the blastomere isolation.
Tygon S3 Laboratory Tubing,, Formulation E-3603, Inner diameter 3.175 mm Saint Gobain Performance Plastics 89403-862 For the blastomere isolation.
Tygon S3 Laboratory Tubing,, Formulation E-3603, Inner diameter 6.35 mm Saint Gobain Performance Plastics 89403-854 For the blastomere isolation.

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Cite This Article
C. elegans Blastomere Dissection: A Method to Remove the Eggshell and Dissociate Embryonic Cells. J. Vis. Exp. (Pending Publication), e20158, doi: (2023).

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