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Related Experiment Video

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A Simple Method for Generating Cerebral Organoids from Human Pluripotent Stem Cells.

Yean Ju Hong1, So Been Lee1, Joonhyuk Choi1

  • 1Department of Stem Cell and Regenerative Biotechnology, KU Institute of Science and Technology, Konkuk University, Seoul, Korea.

International Journal of Stem Cells
|February 27, 2022
PubMed
Summary

Researchers optimized a method for generating cerebral organoids from human pluripotent stem cells. This protocol ensures consistent production of neural progenitor cells and neurons for neural differentiation research.

Keywords:
Brain organoidDifferentiationHuman pluripotent stem cellsOrganoidPluripotency

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Area of Science:

  • Neuroscience
  • Stem Cell Biology
  • Developmental Biology

Background:

  • Brain organoid technology represents a significant advancement in neural differentiation research.
  • Optimized methods for establishing cerebral organoids from human pluripotent stem cells are crucial.

Purpose of the Study:

  • To optimize a feeder-free method for generating cerebral organoids from human pluripotent stem cells.
  • To ensure consistent and robust production of cerebral organoids containing neural progenitor cells and neurons.
  • To develop a protocol applicable to both human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs).

Main Methods:

  • Investigated the morphological changes during cerebral organoid differentiation, including neuroepithelium, neural tube, and neural folding.
  • Characterized cerebral organoids using immunocytochemical staining on Cryostat and Vibratome-prepared sections.
  • Established a routine method for generating early cerebral organoids with a cortical layer and neural progenitor zone.

Main Results:

  • Successfully optimized a method for cerebral organoid generation from feeder-free cultured human pluripotent stem cells.
  • Demonstrated consistent and robust production of cerebral organoids comprising neural progenitor cells and neurons.
  • Established a routine method for generating early cerebral organoids with distinct cortical layers and neural progenitor zones.

Conclusions:

  • An optimized methodology for cerebral organoid generation using hESCs and hiPSCs was developed.
  • The protocol yields consistent and efficient cerebral organoids from both hESCs and hiPSCs.
  • Morphological analysis of brain organoids can be performed using 2D (immunostaining/sectioning) or 3D (whole tissue staining/clarification) methods.