Home › Cell Biology › Dissection, Culture and Analysis of Primary Cranial Neural Crest Cells from Mouse for the Study of Neural Crest Cell Delamination and Migration
Cell Biology JoVE (Open Access) Citable · DOI

Dissection, Culture and Analysis of Primary Cranial Neural Crest Cells from Mouse for the Study of Neural Crest Cell Delamination and Migration

DOI: 10.3791/60051-v
What you'll learn
  • ✓ Dissect and culture primary cranial neural crest cells from mouse embryos
  • ✓ Perform live-cell imaging and single-cell tracking of migrating neural crest cells
  • ✓ Quantify cell migration speed, morphology, and shape dynamics from time-lapse data
Protocol

This protocol describes the dissection and culture of cranial neural crest cells from mouse models, primarily for the study of cell migration. We describe the live imaging techniques used and the analysis of speed and cell shape changes.

Difficulty
advanced
Total time
~2 days (Day 1: dissection and culture initiation; Day 2: live imaging and analysis)
Model organism
Mouse (early somite stage embryos)
Biosafety
BSL-1

Steps

1
Dissect early somite stage mouse embryos

Isolate cranial neural crest tissue from mouse embryos at the early somite stage. Prepare tissue for culture by careful microdissection under a stereomicroscope.

▶ 01:05
2
Perform live-cell imaging of neural crest cultures

Set up time-lapse microscopy imaging of murine cranial neural crest cells in culture. Acquire image sequences capturing cell migration and morphological changes over time.

▶ 03:12
3
Track single neural crest cells across frames

Manually or computationally identify and follow individual cell trajectories throughout the time-lapse sequence to quantify migration paths.

▶ 04:36
4
Quantify cell migration speed and displacement

Measure migration velocity and total displacement distance for tracked cells using image analysis software to determine migratory capacity.

▶ 05:22
5
Measure cell area and circularity metrics

Quantify cell morphology parameters including total cell area and circularity index from segmented cell images to assess shape dynamics during migration.

▶ 06:04
6
Analyze representative results and cell dynamics

Review quantified data on migration speed, trajectory, cell area, and shape changes. Present example traces and morphological measurements demonstrating protocol outcomes.

▶ 07:00
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