C. elegans Dissection blastomere: Une méthode pour enlever la coquille d’œuf et dissocier les cellules embryonnaires
Published: April 30, 2023
Abstract
Source : Hsu, C. R., et autres. Reconstitution in vitro des modèles de contact cellulaire spatial avec des blastomères embryonnaires isolés de Caenorhabditis elegans et des perles de polystyrène adhésif. J. Vis. Exp. (2019).
Cette vidéo introduit une méthode d’isolement des blastomères C. elegans à partir d’embryons précoces. Les cellules qui en résultent conviennent à la culture cellulaire ou à des expériences ex vivo.
Protocol
Ce protocole est un extrait de Hsu et coll., In Vitro Reconstitution of Spatial Cell Contact Patterns with Isolated Caenorhabditis elegans Embryo Blastomeres and Adhesive Polystyrene Beads, J. Vis. Exp. (2019). Isolement du blastomere embryonnaire Portez des gants et un manteau de laboratoire pour éviter les coupures et l…
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. |
Citar este artigo
C. elegans Blastomere Dissection: A Method to Remove the Eggshell and Dissociate Embryonic Cells. J. Vis. Exp. (Pending Publication), e20158, doi: (2023).