Engineering induction of singular neural rosette emergence within hPSC-derived tissues

Area of Science:

• Stem cell biology
• Developmental neuroscience
• Biophysics

Background:

• Human pluripotent stem cell (hPSC)-derived neural organoids offer insights into central nervous system (CNS) development.
• Current organoid models generate multiple independent neural rosettes, hindering coordinated development.
• A singular neuroepithelial tube is the in vivo precursor to the entire CNS.

Purpose of the Study:

• To investigate the role of initial tissue morphology in controlling neural rosette formation in hPSC-derived neural organoids.
• To determine if controlling morphology can induce singular neural rosette development for reproducible CNS tissue biomanufacture.

Main Methods:

• hPSC culture and neural differentiation protocols.
• Controlled geometric confinement to dictate initial tissue morphology.
• ROCK signaling pathway inhibition and assessment of cell contractility.
• Microscopy and imaging to analyze neural rosette formation and tissue development.

Main Results:

• Controlling initial tissue morphology successfully induced single neural rosette emergence in >80% of hPSC-derived forebrain and spinal tissues.
• Optimal morphology for singular rosette induction differed between forebrain and spinal tissues, linked to ROCK-mediated contractility.
• Released tissues exhibited radial outgrowth, maintaining a singular neuroepithelium and peripheral neuronal differentiation.

Conclusions:

• Neural tissue morphology is a critical biophysical parameter for controlling neural organoid development.
• This finding advances the biomanufacture of CNS tissues with biomimetic anatomy and physiology.
• Targeting morphology offers a strategy for reproducible in vitro neural tissue development.