C. 예인대 Blastomere 해부 : 달걀 껍질을 제거하고 배아 세포를 해리하는 방법

Published: April 30, 2023

Abstract

출처: Hsu, C. R., . 고립 된 Caenorhabditis elegans 배아 Blastomeres 및 접착 폴리스티렌 구슬과 공간 세포 접촉 패턴의 시험관 재구성. J. 비스 익스펙. (2019). 

이 비디오는 초기 배아에서 C. elegans blastomeres를 격리하는 방법을 소개합니다.  결과 세포는 세포 배양 또는 전 생체 내 실험에 적합합니다.

Protocol

이 프로토콜은 Hsu 외에서 발췌,   격리 된 Caenorhabditis elegans 배아 Blastomeres 및 접착 폴리스티렌 구슬, J. Vis. Exp. (2019)와 공간 세포 접촉 패턴의생체로 재구성이다. 배아 블라도미어의 격리 장갑과 실험실 코트를 착용하여 절단및 표백 용액과 접촉하?…

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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