Deciphering brain organoids heterogeneity by identifying key quality determinants

organoids
brain
rna-seq
Authors

Daniil Kachkin

Naime Zagha

Tom Boerstler

Federica Furlanetto

Negar Nayebzade

Luke Zappia

Michelle Boisvert

Michaela Farrell

Sonja Ploetz

Martin Regensburger

Claudia Günther

Juergen Winkler

Pooja Gupta

Fabian Theis

Marisa Karow

Sven Falk

Beate Winner

Florian Krach

Date

January 13, 2025

Links
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Abstract

Brain organoids derived from human pluripotent stem cells (hPSCs) hold immense potential for modeling neurodevelopmental processes and disorders. However, their experimental variability and undefined organoid selection criteria for analysis hinder reproducibility. As part of the Bavarian ForInter consortium, we generated 72 brain organoids from distinct hPSC lines. We conducted a comprehensive analysis of their morphological and cellular characteristics at an early stage of their development. In our assessment, the Feret diameter emerged as a reliable, single parameter that characterizes brain organoid quality. Transcriptomic analysis further confirmed the reliability of this marker and identified a negative impact of mesenchymal cells on the abundance of organoid formation. High-quality organoids consistently displayed a lower mesenchymal cell presence. These findings offer a framework for enhancing brain organoid standardization and reproducibility, underscoring the need for morphological quality controls and the consideration of mesenchymal cell influence on organoid-based modeling.

Citation

BibTeX citation:
@misc{kachkin2025,
  author = {Kachkin, Daniil and Zagha, Naime and Boerstler, Tom and
    Furlanetto, Federica and Nayebzade, Negar and Zappia, Luke and
    Boisvert, Michelle and Farrell, Michaela and Ploetz, Sonja and
    Regensburger, Martin and Günther, Claudia and Winkler, Juergen and
    Gupta, Pooja and Theis, Fabian and Karow, Marisa and Falk, Sven and
    Winner, Beate and Krach, Florian},
  title = {Deciphering Brain Organoids Heterogeneity by Identifying Key
    Quality Determinants},
  date = {2025-01-13},
  url = {https://doi.org/10.1101/2025.01.13.632763},
  doi = {10.1101/2025.01.13.632763},
  langid = {en},
  abstract = {Brain organoids derived from human pluripotent stem cells
    (hPSCs) hold immense potential for modeling neurodevelopmental
    processes and disorders. However, their experimental variability and
    undefined organoid selection criteria for analysis hinder
    reproducibility. As part of the Bavarian ForInter consortium, we
    generated 72 brain organoids from distinct hPSC lines. We conducted
    a comprehensive analysis of their morphological and cellular
    characteristics at an early stage of their development. In our
    assessment, the Feret diameter emerged as a reliable, single
    parameter that characterizes brain organoid quality. Transcriptomic
    analysis further confirmed the reliability of this marker and
    identified a negative impact of mesenchymal cells on the abundance
    of organoid formation. High-quality organoids consistently displayed
    a lower mesenchymal cell presence. These findings offer a framework
    for enhancing brain organoid standardization and reproducibility,
    underscoring the need for morphological quality controls and the
    consideration of mesenchymal cell influence on organoid-based
    modeling.}
}
For attribution, please cite this work as:
Kachkin, D., Zagha, N., Boerstler, T., Furlanetto, F., Nayebzade, N., Zappia, L., Boisvert, M., Farrell, M., Ploetz, S., Regensburger, M., Günther, C., Winkler, J., Gupta, P., Theis, F., Karow, M., Falk, S., Winner, B. & Krach, F. Deciphering brain organoids heterogeneity by identifying key quality determinants. bioRxiv Preprint at https://doi.org/10.1101/2025.01.13.632763 (2025)