Neuroscience JoVE (Open Access) Citable · DOI

An Explant Assay for Assessing Cellular Behavior of the Cranial Mesenchyme

Name: An Explant Assay for Assessing Cellular Behavior of the Cranial Mesenchyme
Uploaded: 2026-06-17
Description: The central nervous system is derived from the neural plate that undergoes a series of complex morphogenetic movements resulting in formation of the ...

DOI: 10.3791/4245-v

What you'll learn

✓ Isolate and prepare cranial mesenchyme explants from embryonic tissue
✓ Observe and characterize cellular migration behavior in ex vivo culture
✓ Apply pharmacological manipulations to study mesenchymal cell dynamics
✓ Perform live imaging analysis of neural fold morphogenesis

Protocol

The cranial mesenchyme undergoes dramatic morphogenic movements that likely provides a driving force for elevation of the neural folds1,2. Here we describe a simple ex vivo explant assay to characterize the cellular behaviors of the cranial mesenchyme during neurulation. This assay has numerous applications including being amenable to pharmacological manipulations and live imaging analyses.

Difficulty

intermediate

Total time

~3–4 hours per experiment (embryo collection, dissection, and culture setup)

Model organism

Xenopus laevis

Biosafety

BSL-1

Steps

Prepare equipment and reagents for explant culture

Set up the dissection microscope, culture dishes, and necessary buffered solutions. Ensure all materials are sterile and maintained at appropriate temperature for embryo handling.

▶ 01:33

Collect and stage embryonic specimens

Obtain embryos at the appropriate developmental stage (typically early neurula). Sort by morphological criteria to ensure uniform developmental progression.

▶ 02:56

Isolate cranial mesenchyme tissue explants

Carefully dissect the cranial mesenchymal region from intact embryos using fine forceps and microscopy. Transfer isolated explants to culture medium for subsequent analysis.

▶ 04:38

Assess cell migration from explant tissue

Culture explants and observe outward migration of mesenchymal cells over time. Document morphology and movement patterns using phase-contrast or live fluorescence imaging.

▶ 06:49

💬 Comments coming soon