Transcriptome profiling of human pluripotent stem cell-derived cerebellar organoids reveals faster commitment under dynamic conditions

Teresa P. Silva, Rui Sousa-Luís, Tiago G. Fernandes, Evguenia P. Bekman, Carlos A.V. Rodrigues, Sandra H. Vaz, Leonilde M. Moreira, Yas Hashimura, Sunghoon Jung, Brian Lee, Maria Carmo-Fonseca, Joaquim M.S. Cabral

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Human-induced pluripotent stem cells (iPSCs) have great potential for disease modeling. However, generating iPSC-derived models to study brain diseases remains a challenge. In particular, the ability to recapitulate cerebellar development in vitro is still limited. We presented a reproducible and scalable production of cerebellar organoids by using the novel single-use Vertical-Wheel bioreactors, in which functional cerebellar neurons were obtained. Here, we evaluate the global gene expression profiles by RNA sequencing (RNA-seq) across cerebellar differentiation, demonstrating a faster cerebellar commitment in this novel dynamic differentiation protocol. Furthermore, transcriptomic profiles suggest a significant enrichment of extracellular matrix (ECM) in dynamic-derived cerebellar organoids, which can better mimic the neural microenvironment and support a consistent neuronal network. Thus, an efficient generation of organoids with cerebellar identity was achieved for the first time in a continuous process using a dynamic system without the need of organoids encapsulation in ECM-based hydrogels, allowing the possibility of large-scale production and application in high-throughput processes. The presence of factors that favors angiogenesis onset was also detected in dynamic conditions, which can enhance functional maturation of cerebellar organoids. We anticipate that large-scale production of cerebellar organoids may help developing models for drug screening, toxicological tests, and studying pathological pathways involved in cerebellar degeneration.

Original languageEnglish
Pages (from-to)2781-2803
Number of pages23
JournalBiotechnology and Bioengineering
Volume118
Issue number7
DOIs
Publication statusPublished - Jul 2021
Externally publishedYes

Keywords

  • cerebellum
  • dynamic conditions
  • human pluripotent stem cells
  • large-scale production
  • organoids

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