Transfection into Mouse/Rat Cultured Embryos by Ex Utero Electroporation

Transfection into Mouse/Rat Cultured Embryos by Ex Utero Electroporation

APPLICATIONS

Electroporation for mammalian embryos in the whole embryo culture system

Procedures

  1. ProcedureProcedure1: Pre-culture embryos in the whole embryo culture system for 1.5-2 hours prior to electroporation.
  2. Place the embryo in a Petri dish with Tyrode’s solution.
  3. Inject 0.1-0.5 μl of plasmid DNA into the brain ventricle with a fine capillary.
  4. Apply square pulses using the electroporator (CUY21) and electrodes (chamber-type and forceps-type electrodes are available). 70V, 50 msec at 1-second interval, five pulses are applied for E10.5 mouse embryos.
  5. Culture the electroporated embryos for 24-48 hours in the whole embryo culture system.Transfection of a fluorescent protein-expression vector into the developing rat cortex.

Transfection of a fluorescent protein-expression vector into the developing rat cortex.

(A) EGFP-expression vector was electroporated into E11.5 rat telencephalon. The electroporated embryo was cultured in the whole embryo culture system (WEC).

(B) 24 hours after electroporation, EGFP-expression was specifically detected at the dorsal part of the telencephalon.

Tel: telencephalon;
Di: diencephalon

Transfection of fluorescent protein-expression vectors into the rat spinal cord.

(A) EGFP-expression vector was transfected into the spinal cord of E12.5 rat embryos by electroporation. The electroporated embryos were cultured for 24 hours in the whole embryo culture system (WEC).

(B-E) Time-lapse analysis of neuroepithelial cells in the slice culture system. Histon-EGFP- and DsRed2-expression vectors were co-electroporated into the E12.0 rat spinal cord. The electroporated embryo was cultured for 24 hours in the WEC, then the spinal cord was sliced and time-lapse recording was performed.
(B-E) Cell nuclei (B, green) and cytoplasms (C, magenta) of neuroepithelial cells in the slice are simultaneously labeled by the electroporation
(E). (D) A DIC image.
hb: hindbrain; sc: spinal cord; fl: forelimb; DIC: differential interference contrast.

Masanori Takahashi, Tadashi Nomura, Noriko Osumi, Department of Developmental Neurobiology, Tohoku University Graduate School of Medicine
*Differentiation, Volume 70, Issue 4-5, Pages 155-162, June 2002

PUBLICATIONS

Electroporation

Drug Delivery and Transfection

Electro Cell Fusion

Fluorescent Staining

Single-Cell/Micro-Particle Transfer

Cell Freezing

Mechanical Vibration