Spinal Cord Neuron Isolation: A Density Gradient Procedure to Isolate Neonatal Spinal Cord Neurons for In Vitro Studies

Abstract

Source: Eldeiry, M. et al. Spinal Cord Neurons Isolation and Culture from Neonatal Mice. J. Vis. Exp. (2017)

In this video, we describe a step-by-step protocol for isolating spinal cord neurons from a neonatal mouse for in vitro studies.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Harvesting Spinal Cords

NOTE: All instruments should be autoclaved (135 °C and 30 psi for 4 x 7 min cycles) for sterility.

1. Euthanize 1–3 day-old C57BL/6 mouse pups in a chamber with isoflurane. Wait 30 s after the cessation of movement and pinch the leg to confirm a lack of response.
2. Separate the head from the body using scissors, with pup in the prone position.
3. Stabilize the hind legs or tai....