Human cortical spheroids with a high diversity of innately developing brain cell types

Area of Science:

• Neuroscience
• Developmental Biology
• Stem Cell Biology

Background:

• Three-dimensional (3D) human brain spheroids are crucial for studying central nervous system (CNS) development and function.
• Current models lack cell-type diversity, limiting integrated exploration of CNS disease mechanisms.

Purpose of the Study:

• To develop a reproducible 5-month culture protocol for human cortical spheroids (hCSs) with a broad range of cell types.
• To establish a versatile 3D human brain cell model for studying intercellular CNS communication and neurological diseases.

Main Methods:

• Utilized H9 embryonic stem cells for spheroid generation.
• Employed a 5-month culture protocol.
• Validated cell types using RNA-sequencing, qPCR, immunocytochemistry, and transmission electron microscopy.

Main Results:

• Generated hCSs containing neuroectoderm-derived neural progenitors, mature neurons, astrocytes, and oligodendrocyte precursor cells.
• Confirmed the presence of mesoderm-derived microglia and endothelial cells.
• Transcriptomic analysis showed hCSs resemble fetal human brain tissue (19-26 weeks gestational age).
• Demonstrated a neuroinflammatory response upon pro-inflammatory stimulation, proving model applicability.

Conclusions:

• Developed a 3D human brain cell model with diverse, innately developing neuroectoderm- and mesoderm-derived cell types.
• The protocol provides a versatile platform for comprehensive examination of CNS intercellular communication.
• This model facilitates advanced study of neurological disease mechanisms.