Generation of spatially patterned human neural tube-like structures using microfluidic gradient devices.

Area of Science:

• Developmental Biology
• Stem Cell Biology
• Bioengineering

Background:

• Human neural tube formation is crucial for nervous system development.
• Human pluripotent stem cells (hPSCs) offer a valuable model for studying neurodevelopment.
• Traditional animal models have limitations in recapitulating human neural development.

Purpose of the Study:

• To design and implement a microfluidic gradient device for modeling human neural tube formation and regional patterning.
• To create microfluidic neural tube-like structures (μNTLS) and forebrain-like structures (μFBLS) using hPSCs.
• To establish a robust system for studying human neurodevelopment and related diseases.

Main Methods:

• Utilized a microfluidic gradient device to control chemical gradients for patterning hPSCs.
• Formed tubular or spherical colonies of hPSCs within microfluidic channels.
• Exposed cell colonies to controlled gradients for rostral-caudal and dorsal-ventral patterning.

Main Results:

• Successfully generated μNTLS and μFBLS that mimic early human neural development.
• μNTLS exhibited lumenal structures, regional marker expression, and neural crest development.
• μFBLS showed segregated dorsal/ventral regions and layered neuronal development, mirroring pallium/subpallium formation.
• Both structures supported long-term culture, imaging, and molecular analyses.

Conclusions:

• The microfluidic system provides a powerful platform for studying human neurodevelopment and disease modeling.
• The developed μNTLS and μFBLS recapitulate critical aspects of early human neural patterning.
• This protocol is adaptable for researchers with expertise in soft lithography and cell culture.