(T1090) SPINAL CORD ORGANOIDS (SCOS) ARE VALUABLE MODELS FOR DEVELOPMENT AND DISEASES. WE PERFORMED SINGLE-CELL RNA-SEQ ON SCOS (D30–120), REVEALING DIVERSE CELL TYPES AND A DETAILED MOLECULAR ATLAS OVER TIME.

Abstract: Stem cell-derived organoids are valuable models that recapitulate human physiology in remarkable detail and offer unique opportunities to study central nervous system diseases that are challenging to model using animals or monolayer cultures. Our lab has established a robust protocol for generating spinal cord organoids (SCOs) from induced pluripotent stem cells (iPSCs) to investigate spinal cord development, injury, and neurological disorders. SCOs progress through defined developmental stages: spinal cord progenitor cells (SCPCs) at day 10, motor neurons and interneurons by day 30, and astrocytes and oligodendrocytes by days 60 and 90, as confirmed by immunohistochemistry.
However, A comprehensive whole-transcriptome analysis of SCOs at specific time points has been hindered by technical limitations in dissociating organoids into viable single cells. Furthermore, while SCPC transplantation holds promise as a therapeutic approach for spinal cord injury, the differentiation and integration of these cells in vivo remain poorly understood.
In this study, we developed a novel protocol to efficiently dissociate SCOs into viable single cells, enabling single-cell RNA sequencing (scRNA-SEQ) from day 30 to day 120. Our data provide a detailed molecular atlas and identify diverse cell types within SCOs at these stages. Additionally, we performed scRNA-SEQ on dissociated rat spinal cords following human SCPC transplantation, obtaining both rat and human cells. By mapping reads to a combined rat-human genome, we observed that cell types derived from day-60 organoids closely resemble human cell types differentiated in vivo after SCPC transplantation. This finding highlights the utility of SCOs as predictive models for the regenerative potential of transplanted cells.
Our work establishes a comprehensive molecular and cellular framework for spinal cord organoids, offering valuable insights into spinal cord biology and advancing their application in regenerative medicine and disease modelling.

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Clinical Trial ID number: N.A.