Neuroscience JoVE (Open Access) Citable · DOI

The Culture of Primary Motor and Sensory Neurons in Defined Media on Electrospun Poly-L-lactide Nanofiber Scaffolds

Name: The Culture of Primary Motor and Sensory Neurons in Defined Media on Electrospun Poly-L-lactide Nanofiber Scaffolds
Uploaded: 2026-07-01
Duration: 16 min 3 s
Description: Electrospinning is a technique for producing micro- to nano-scale fibers. Fibers can be electrospun with varying degrees of alignment, from highly aligned ...

DOI: 10.3791/2389-v

What you'll learn

✓ Prepare electrospun poly-L-lactide nanofiber scaffolds for neuron culture
✓ Culture primary rat embryonic motor and sensory neurons in serum-free media
✓ Identify and visualize neurons using immunocytochemistry on aligned fiber substrates

Protocol

Aligned electrospun fibers direct the growth of neurons in vitro and are a potential component of nerve regeneration scaffolds. We describe a procedure for preparing electrospun fiber substrates and the serum-free culture of primary rat E15 sensory (DRG) and motor neurons. Visualization of neurons by immunocytochemistry is also included.

Difficulty

advanced

Total time

~4–5 days (including scaffold preparation, embryo dissection, cell isolation, and culture maturation)

Model organism

Rat (Rattus norvegicus), embryonic day 15 (E15)

Biosafety

BSL-1

Steps

Prepare electrospun fiber substrates

Fabricate aligned electrospun poly-L-lactide nanofiber scaffolds using electrospinning equipment. Treat and sterilize substrates for cell culture use.

▶ 01:43

Isolate primary rat embryonic neurons

Harvest embryos at E15 and dissect brain/spinal cord tissue for primary motor and sensory neuron extraction.

▶ 04:40

Process motor neurons from tissue

Enzymatically dissociate and mechanically tritturate embryonic motor neuron tissue to generate single-cell suspension.

▶ 06:53

Process sensory neurons from DRG

Enzymatically dissociate and mechanically tritturate dorsal root ganglion (DRG) tissue to generate sensory neuron single-cell suspension.

▶ 09:29

Plate neurons and culture in defined media

Seed motor and sensory neurons onto electrospun fiber substrates at defined density in serum-free culture medium. Maintain in vitro culture conditions.

▶ 10:56

Visualize neurons by immunocytochemistry

Fix cultured neurons and perform immunostaining to identify motor (e.g., choline acetyltransferase) and sensory (e.g., neuronal markers) neuron populations.

▶ 12:41

💬 Comments coming soon