A framework for neural organoids, assembloids and transplantation studies

Sergiu P Pașca^{1

2}, Paola Arlotta^{3

4}, Helen S Bateup^{5

6}, J Gray Camp^{7

8}, Silvia Cappello^{9

10}, Fred H Gage¹¹, Jürgen A Knoblich^{12

13}, Arnold R Kriegstein^{14

15}, Madeline A Lancaster^{16

17}, Guo-Li Ming^{18

19

20

21}, Gaia Novarino²², Hideyuki Okano²³, Malin Parmar²⁴, In-Hyun Park^{25

26}, Orly Reiner^{27

28}, Hongjun Song^{18

19

20

29}, Lorenz Studer³⁰, Jun Takahashi³¹, Sally Temple³², Giuseppe Testa^{33

34}, Barbara Treutlein³⁵, Flora M Vaccarino^{26

36

37

38}, Pierre Vanderhaeghen^{39

40}, Tracy Young-Pearse^{41

42}

Affiliations

¹ Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA. spasca@stanford.edu.
² Stanford Brain Organogenesis, Wu Tsai Neurosciences Institute and Bio-X, Stanford University, Stanford, CA, USA. spasca@stanford.edu.
³ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
⁴ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁵ Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, USA.
⁶ Department of Neuroscience, University of California Berkeley, Berkeley, CA, USA.
⁷ Institute of Human Biology (IHB), Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel, Switzerland.
⁸ Biozentrum, University of Basel, Basel, Switzerland.
⁹ Department of Physiological Genomics, Biomedical Center (BMC), Faculty of Medicine, Ludwig Maximilian University of Munich, Munich, Germany.
¹⁰ Max Planck Institute of Psychiatry, Munich, Germany.
¹¹ Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
¹² Institute of Molecular Biotechnology, Austrian Academy of Sciences, Vienna Biocenter, Vienna, Austria.
¹³ Department of Neurology, Medical University of Vienna, Vienna, Austria.
¹⁴ Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA.
¹⁵ Department of Neurology, University of California San Francisco, San Francisco, CA, USA.
¹⁶ MRC Laboratory of Molecular Biology, Cambridge, UK.
¹⁷ Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.
¹⁸ Department of Neuroscience and Mahoney Institute for Neurosciences, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹⁹ Department of Cell and Developmental Biology, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
²⁰ Institute for Regenerative Medicine, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
²¹ Department of Psychiatry, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
²² Institute of Science and Technology of Austria, Klosterneuburg, Austria.
²³ Keio University Regenerative Medicine Research Center, Kanagawa, Japan.
²⁴ Department of Experimental Medical Science, Lund Stem Cell Center, Lund University, Lund, Sweden.
²⁵ Department of Genetics, Yale School of Medicine, New Haven, CT, USA.
²⁶ Yale Stem Cell Center, Yale University, New Haven, CT, USA.
²⁷ Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
²⁸ Department of Molecular Neuroscience, Weizmann Institute of Science, Rehovot, Israel.
²⁹ The Epigenetics Institute, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
³⁰ The Center for Stem Cell Biology, Developmental Biology Program, Sloan Kettering Institute for Cancer Research, New York, NY, USA.
³¹ Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
³² Neural Stem Cell Institute, Albany, NY, USA.
³³ Department of Oncology and Hemato-Oncology, University of Milan, Milan, Italy.
³⁴ Human Technopole, Viale Rita Levi Montalcini, Milan, Italy.
³⁵ Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland.
³⁶ Child Study Center, Yale University, New Haven, CT, USA.
³⁷ Department of Neuroscience, Yale University, New Haven, CT, USA.
³⁸ Yale Kavli Institute for Neuroscience, New Haven, CT, USA.
³⁹ VIB-KU Leuven Center for Brain and Disease Research, Leuven, Belgium.
⁴⁰ Department of Neurosciences, Leuven Brain Institute, Leuven, Belgium.
⁴¹ Department of Neurology, Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.
⁴² Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA.

PMID: 39653126
DOI: 10.1038/s41586-024-08487-6

A framework for neural organoids, assembloids and transplantation studies

Sergiu P Pașca et al. Nature. 2025 Mar.

. 2025 Mar;639(8054):315-320.

doi: 10.1038/s41586-024-08487-6. Epub 2024 Dec 9.

Authors

Affiliations

¹ Department of Psychiatry and Behavioral Sciences, Stanford University, Stanford, CA, USA. spasca@stanford.edu.
² Stanford Brain Organogenesis, Wu Tsai Neurosciences Institute and Bio-X, Stanford University, Stanford, CA, USA. spasca@stanford.edu.
³ Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
⁴ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁵ Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, CA, USA.
⁶ Department of Neuroscience, University of California Berkeley, Berkeley, CA, USA.
⁷ Institute of Human Biology (IHB), Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel, Switzerland.
⁸ Biozentrum, University of Basel, Basel, Switzerland.
⁹ Department of Physiological Genomics, Biomedical Center (BMC), Faculty of Medicine, Ludwig Maximilian University of Munich, Munich, Germany.
¹⁰ Max Planck Institute of Psychiatry, Munich, Germany.
¹¹ Laboratory of Genetics, The Salk Institute for Biological Studies, La Jolla, CA, USA.
¹² Institute of Molecular Biotechnology, Austrian Academy of Sciences, Vienna Biocenter, Vienna, Austria.
¹³ Department of Neurology, Medical University of Vienna, Vienna, Austria.
¹⁴ Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA.
¹⁵ Department of Neurology, University of California San Francisco, San Francisco, CA, USA.
¹⁶ MRC Laboratory of Molecular Biology, Cambridge, UK.
¹⁷ Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.
¹⁸ Department of Neuroscience and Mahoney Institute for Neurosciences, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
¹⁹ Department of Cell and Developmental Biology, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
²⁰ Institute for Regenerative Medicine, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
²¹ Department of Psychiatry, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
²² Institute of Science and Technology of Austria, Klosterneuburg, Austria.
²³ Keio University Regenerative Medicine Research Center, Kanagawa, Japan.
²⁴ Department of Experimental Medical Science, Lund Stem Cell Center, Lund University, Lund, Sweden.
²⁵ Department of Genetics, Yale School of Medicine, New Haven, CT, USA.
²⁶ Yale Stem Cell Center, Yale University, New Haven, CT, USA.
²⁷ Department of Molecular Genetics, Weizmann Institute of Science, Rehovot, Israel.
²⁸ Department of Molecular Neuroscience, Weizmann Institute of Science, Rehovot, Israel.
²⁹ The Epigenetics Institute, Perelman School for Medicine, University of Pennsylvania, Philadelphia, PA, USA.
³⁰ The Center for Stem Cell Biology, Developmental Biology Program, Sloan Kettering Institute for Cancer Research, New York, NY, USA.
³¹ Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
³² Neural Stem Cell Institute, Albany, NY, USA.
³³ Department of Oncology and Hemato-Oncology, University of Milan, Milan, Italy.
³⁴ Human Technopole, Viale Rita Levi Montalcini, Milan, Italy.
³⁵ Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland.
³⁶ Child Study Center, Yale University, New Haven, CT, USA.
³⁷ Department of Neuroscience, Yale University, New Haven, CT, USA.
³⁸ Yale Kavli Institute for Neuroscience, New Haven, CT, USA.
³⁹ VIB-KU Leuven Center for Brain and Disease Research, Leuven, Belgium.
⁴⁰ Department of Neurosciences, Leuven Brain Institute, Leuven, Belgium.
⁴¹ Department of Neurology, Ann Romney Center for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.
⁴² Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA.

PMID: 39653126
DOI: 10.1038/s41586-024-08487-6

Abstract

As the field of neural organoids and assembloids expands, there is an emergent need for guidance and advice on designing, conducting and reporting experiments to increase the reproducibility and utility of these models. In this Perspective, we present a framework for the experimental process that encompasses ensuring the quality and integrity of human pluripotent stem cells, characterizing and manipulating neural cells in vitro, transplantation techniques and considerations for modelling human development, evolution and disease. As with all scientific endeavours, we advocate for rigorous experimental designs tailored to explicit scientific questions as well as transparent methodologies and data sharing to provide useful knowledge for current research practices and for developing regulatory standards.

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Conflict of interest statement

Competing interests: The authors declare no competing interests.

References

1. Pasca, S. P. et al. A nomenclature consensus for nervous system organoids and assembloids. Nature 609, 907–910 (2022). This consensus article provided a framework for the nomenclature of human multicellular models of nervous system development and disease. - DOI - PubMed - PMC
1. The RIVER working group. Reporting in vitro experiments responsibly—the RIVER recommendations. Preprint at MetaArxiv https://doi.org/10.31222/osf.io/x6aut (2023).
1. Lezmi, E., Jung, J. & Benvenisty, N. High prevalence of acquired cancer-related mutations in 146 human pluripotent stem cell lines and their differentiated derivatives. Nat. Biotechnol. 42, 1667–1671 (2024). - DOI - PubMed
1. Panda, A. et al. Clonally selected lines after CRISPR–Cas editing are not isogenic. CRISPR J. 6, 176–182 (2023). - DOI - PubMed - PMC
1. Abyzov, A. et al. Somatic copy number mosaicism in human skin revealed by induced pluripotent stem cells. Nature 492, 438–442 (2012). - DOI - PubMed - PMC

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A framework for neural organoids, assembloids and transplantation studies

Affiliations

A framework for neural organoids, assembloids and transplantation studies

Authors

Affiliations

Abstract

Conflict of interest statement

References

MeSH terms

Grants and funding

LinkOut - more resources

Full Text Sources

Research Materials