Metabolic Assessment in Human Pluripotent Stem Cell-Derived Cerebral Organoids Using HR-MAS NMR Spectroscopy.

Area of Science:

• Neuroscience
• Biochemistry
• Stem Cell Biology

Background:

• Brain metabolism is crucial for function and altered in disease.
• Studying brain metabolism directly in humans is difficult.
• Animal models face welfare and validity concerns, necessitating alternative methods.

Purpose of the Study:

• To establish a pipeline for studying neurometabolic pathways in human pluripotent stem cell-derived cerebral organoids (hPSC-COs).
• To characterize the metabolic profile of hPSC-COs using high-resolution magic-angle spinning proton nuclear magnetic resonance (HR-MAS 1H-NMR) spectroscopy.
• To compare the metabolic profile of hPSC-COs with human fetal and adult brain tissue.

Main Methods:

• Application of HR-MAS 1H-NMR spectroscopy to intact hPSC-COs.
• Quantification of 17 distinct metabolites across various developmental stages of COs.
• Analysis of spectral quality, including linewidth and signal-to-noise ratio (SNR), for metabolite detection.

Main Results:

• Successfully identified and quantified 17 metabolites in hPSC-COs aged 85 to 312 days.
• Detected high spectral quality (linewidth < 4 Hz, SNR > 65) enabling detailed metabolic analysis.
• Revealed a metabolic profile in hPSC-COs resembling human fetal brain, distinct from adult brain, with elevated lactate and hypotaurine, equimolar glutamate/glutamine, and low N-acetylaspartate.

Conclusions:

• HR-MAS 1H-NMR spectroscopy provides direct metabolic assessment in intact hPSC-COs.
• This methodology establishes a valuable platform for investigating human brain metabolism.
• The study offers a novel approach for modeling neurometabolic alterations in neurodegenerative diseases using engineered human brain tissues.