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Related Concept Videos

Organization of the Brain01:30

Organization of the Brain

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The brain is an integral component of the nervous system and serves as the center for processing sensory inputs, making decisions, and directing bodily actions. This complex organ is organized into three primary sections: the hindbrain, midbrain, and forebrain, each responsible for a range of vital functions.
Hindbrain
The hindbrain, located at the base of the brain, plays a vital role in regulating automatic processes that sustain life. It includes the medulla oblongata, which is essential for...
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Related Experiment Video

Updated: Apr 30, 2026

Generation of Standardized and Reproducible Forebrain-type Cerebral Organoids from Human Induced Pluripotent Stem Cells
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Generation of Individualized, Standardized, and Electrically Synchronized Human Midbrain Organoids.

Sanae El Harane1,2, Bahareh Nazari1, Nadia El Harane1,2

  • 1Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, 1206 Geneva, Switzerland.

Cells
|August 13, 2025
PubMed
Summary

Air-liquid interface (ALi) technology, AirLiwell, improves organoid culture by enhancing standardization and cell purity. This method optimizes midbrain organoid development for disease modeling and cell therapy applications.

Keywords:
3D cell cultureAirLiwellParkinson’s diseaseair–liquid interfacecell therapyelectrical recordingsneurospheresorganoidspluripotent stem cellsspheroids

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Area of Science:

  • Biotechnology
  • Developmental Biology
  • Neuroscience

Background:

  • Organoids model human tissues for research but traditional immersion methods face standardization and hypoxia issues.
  • Existing organoid culture methods require high media volumes and agitation, limiting scalability and reproducibility.
  • Issues like organoid fusion and necrosis hinder reliable use in drug discovery and cell therapy.

Purpose of the Study:

  • To develop and validate an air-liquid interface (ALi) culture technology, AirLiwell, for standardized organoid generation.
  • To compare the ALi method with traditional immersion culture using midbrain organoids derived from human pluripotent stem cells (hPSCs).
  • To assess the impact of ALi culture on organoid standardization, cell composition, and functional electrophysiological activity.

Main Methods:

  • Developed AirLiwell, an ALi culture system using non-adhesive microwells for individual organoid maintenance.
  • Cultured midbrain organoids from hPSCs using both ALi and standard immersion methods.
  • Analyzed organoid characteristics using RNA sequencing, immunostaining, single-cell RNA sequencing, and electrophysiology.

Main Results:

  • AirLiwell culture resulted in highly standardized midbrain organoids with optimized size and shape.
  • Single-cell RNA sequencing showed ALi organoids were composed of 99% neural cells (86% neurons), compared to 61% neural cells in immersion cultures.
  • ALi-cultured organoids exhibited synchronized electrophysiological activity, unlike the heterogeneous activity in immersion-cultured organoids.
  • ALi method demonstrated compatibility with Good Manufacturing Practices and versatility for cancer and primary cell organoids.

Conclusions:

  • The AirLiwell ALi technology provides a standardized, scalable platform for organoid culture, improving cell purity and functionality.
  • This method addresses limitations of traditional immersion culture, enhancing reproducibility for applications in developmental biology, drug discovery, and cell therapy.
  • Optimized midbrain organoids are particularly valuable for modeling neurological diseases like Parkinson's and advancing cell-based therapies.