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. |
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).