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Generation of Urine-Derived Induced Pluripotent Stem Cells and Cerebral Organoids for Modeling Down Syndrome

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Abstract

Down syndrome (DS, or trisomy 21, T21), is the most common genetic cause of intellectual disability. Alterations in the complex process of cerebral cortex development contribute to the neurological deficits in DS, although the underlying molecular and cellular mechanisms are not completely understood. Human cerebral organoids (COs) derived from three-dimensional (3D) cultures of induced pluripotent stem cells (iPSCs) provide a new avenue for gaining a better understanding of DS neuropathology. In this study, we aimed to generate iPSCs from individuals with DS (T21-iPSCs) and euploid controls using urine-derived cells, which can be easily and noninvasively obtained from most individuals, and examine their ability to differentiate into neurons and astrocytes grown in monolayer cultures, as well as into 3D COs. We employed nonintegrating episomal vectors to generate urine-derived iPSC lines, and a simple-to-use system to produce COs with forebrain identity. We observed that both T21 and control urine-derived iPSC lines successfully differentiate into neurons and astrocytes in monolayer, as well as into COs that recapitulate early features of human cortical development, including organization of neural progenitor zones, programmed differentiation of excitatory and inhibitory neurons, and upper-and deep-layer cortical neurons as well as astrocytes. Our findings demonstrate for the first time the suitability of using urine-derived iPSC lines to produce COs for modeling DS.

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Data Availability

All data generated or analyzed during this study are available from the corresponding author on request.

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Acknowledgements

We are grateful to all the individuals who participated in this work and their parents/legal guardians. We thank Raquel Vasconcelos Guimarães de Castro, Victor Fernandes Brugnera and Thiago Pinheiro Arrais Aloia for technical assistance. We thank Ana Claudia Pereira Sanguin and Rebeca Passos Costa for administrative assistance.

Funding

This research was funded by Sao Paulo Research Foundation (FAPESP) [A.L.S. grant numbers 2019/08655–2; 2021/14491–2. B.L.Z. grant number: 2018/15371–8]. B.L.Z. was supported by a Young Investigator Fellowship from FAPESP [2019/04859–2].

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Author notes

  1. André Luíz Teles e Silva and Bruno Yukio Yokota contributed equally to this work.

Authors and Affiliations

  1. Hospital Israelita Albert Einstein, São Paulo, SP, Brazil

    André Luíz Teles e Silva, Bruno Yukio Yokota, Andrea Laurato Sertié & Bruna Lancia Zampieri

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  1. André Luíz Teles e Silva
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  2. Bruno Yukio Yokota
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  3. Andrea Laurato Sertié
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  4. Bruna Lancia Zampieri
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Contributions

Conceptualization, A.L.S., B.L.Z.; methodology, A.L.T.S, B.Y.Y., B.L.Z.; formal analysis, A.L.T.S, B.Y.Y., B.L.Z.; writing—original draft preparation, A.L.T.S, B.Y.Y., A.L.S., B.L.Z.; writing—supervision, A.L.S., B.L.Z.; project administration, A.L.S., B.L.Z; funding acquisition, A.L.S., B.L.Z. All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Andrea Laurato Sertié or Bruna Lancia Zampieri.

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Ethics Approval

The study was conducted in accordance with the Declaration of Helsinki, and approved by the Institutional Review Board and Ethics Committee of Hospital Israelita Albert Einstein (CAEE: 06036919.8.0000.0071, 20/02/2019).

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All individuals agreed to participate in the study.

Informed Consent Statement

Written informed consent was obtained from each participant and their parents/legal guardians.

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Not applicable.

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The authors declare no conflict of interest.

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Andrea L. Sertié and Bruna L. Zampieri jointly supervised this work.

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Teles e Silva, A.L., Yokota, B.Y., Sertié, A.L. et al. Generation of Urine-Derived Induced Pluripotent Stem Cells and Cerebral Organoids for Modeling Down Syndrome. Stem Cell Rev and Rep 19, 1116–1123 (2023). https://doi.org/10.1007/s12015-022-10497-8

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