Enrichment of Neural Crest Cells by Antibody Labeling and Flow Cytometry for Single-Cell Transcriptomics in a Lizard

Area of Science:

• Developmental Biology
• Evolutionary Biology
• Cell Biology

Background:

• Neural crest cells (NCCs) are crucial for vertebrate development, contributing to diverse traits.
• Studying NCCs is challenging due to their migratory behavior and heterogeneity.
• Current methods often rely on specific model organisms with advanced genetic tools.

Purpose of the Study:

• To develop a novel, broadly applicable method for isolating NCCs.
• To overcome limitations of existing techniques in non-model organisms.
• To enable transcriptomic profiling of NCCs in a wider range of vertebrates.

Main Methods:

• Antibody labeling combined with fluorescence-activated cell sorting (FACS).
• Application in the common wall lizard (Podarcis muralis).
• Validation using microscopy, RT-qPCR, and single-cell RNA sequencing (scRNA-seq).

Main Results:

• The antibody-FACS method efficiently enriches for NCCs.
• The technique is comparable in efficiency to methods using transgenic animals.
• Transcriptional profiles of NCCs in wall lizard embryos were successfully characterized.

Conclusions:

• This novel method expands the study of NCCs to vertebrates lacking transgenic tools.
• It facilitates deeper understanding of NCC roles in development and evolution.
• Enables comparative transcriptomics of NCCs across diverse species.