NBDC Research ID: hum0472.v1
SUMMARY
Aims: When cerebral cortical neurons are damaged due to stroke or trauma, the injured neurons typically do not recover, resulting in persistent sequelae such as motor paralysis. One potential therapeutic approach is the transplantation of neurons derived from pluripotent stem cells. While we have demonstrated that cerebral organoids can be differentiated from human ES cells, using human iPS cells is more desirable for clinical applications due to the possibility of allogeneic transplantation with HLA compatibility. Here, we will perform non-clinical trials to establish differentiation protocols from hiPSCs to cerebral cortical neurons and assess the safety and efficacy of these differentiated cells. Additionally, we will explore large-scale cell production for future industrialization.
Methods: Organoids were dissociated into single cells with Neuron Dissociation Solutions. Dissociated cells were re-suspended with HBSS supplemented with 10% (v/v) KSR and 10μM Y-27632 at 1,000 cells/µL density. Cell suspension was loaded onto a Chromium Next GEM Chip G (2000177 10X Genomics), targeting 3,000 cells for capture per well, and processed in the Chromium controller to obtain Gel Beads-in-Emulsion. Libraries were generated and RNA-seq was performed.
Participants/Materials: Organoids derived from induced pluripotent stem cells of a single healthy individual.
Dataset ID | Type of Data | Criteria | Release Date |
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JGAS000726 | NGS (scRNA-seq) | Controlled-access (Type I) | 2024/08/23 |
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MOLECULAR DATA
Participants/Materials | Organoids derived from iPS cells of a single healthy individual: 9 samples |
Targets | scRNA-seq |
Target Loci for Capture Methods | - |
Platform | Illumina [NovaSeq 6000] |
Library Source | RNAs extracted from organoids |
Cell Lines | - |
Library Construction (kit name) | Chromium Next GEM Single Cell 3’ Reagent Kits v3.1 |
Fragmentation Methods | enzymatic fragmentation |
Spot Type | Paired-end |
Read Length (without Barcodes, Adaptors, Primers, and Linkers) | 150 bp |
Japanese Genotype-phenotype Archive Dataset ID | JGAD000859 |
Total Data Volume | 401.6 GB (fastq) |
Comments (Policies) | NBDC policy |
DATA PROVIDER
Principal Investigator: Jun Takahashi
Affiliation: Department of Clinical Application, Center for iPS Cell Research and Application, Kyoto University
Project / Group Name: -
URL: https://www.cira.kyoto-u.ac.jp/e/research/takahashi_summary.html
Funds / Grants (Research Project Number):
Name | Title | Project Number |
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Acceleration Program of R&D and Implementation for Regenerative Medicine and Cell and Gene Therapy, Japan Agency for Medical Research and Development (AMED) | Translational study for iPS cell-based therapy for stroke | JP23bm1223005 |
Japan Agency for Medical Research and Development (AMED) | Study of iPS cell-based therapy for stroke | JP24ym0126160 |
PUBLICATIONS
Title | DOI | Dataset ID | |
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1 | Validation of non-invasive morphology-based selection of cerebral cortical organoids by paired morphological and single-cell RNA sequencing analyses | JGAD000859 | |
2 |
USRES (Controlled-access Data)
Principal Investigator | Affiliation | Research Title | Data in Use (Dataset ID) | Period of Data Use |
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