|
| Status |
Public on Nov 24, 2023 |
| Title |
Single-cell transcriptomics and in vitro lineage tracing reveals differential susceptibility of human iPSC-derived midbrain dopaminergic neurons in a cellular model of Parkinson’s disease |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Advances in stem cell technologies open up new avenues for modelling development and diseases. The success of these pursuits however rely on the use of cells most relevant to those targeted by the disease of interest, for example, midbrain dopaminergic neurons for Parkinson’s disease. In the present study, we report the generation of a human induced pluripotent stem cell (iPSC) line capable of purifying and tracing nascent midbrain dopaminergic progenitors and their differentiated progeny via the expression of a Blue Fluorescent Protein (BFP). This was achieved by CRISPR/Cas9 assisted knock-in of BFP and Cre into the safe harbour locus AAVS1 and an early midbrain dopaminergic lineage marker gene LMX1A, respectively. Immunocytochemical analysis and single cell RNA sequencing of iPSC-derived neural cultures confirms developmental recapitulation of the human fetal midbrain and high quality midbrain cells. By modelling Parkinson’s disease-related drug toxicity using 1-Methyl-4-phenylpyridinium (MPP+), we showed preferential reduction of BFP+ cells, a finding demonstrated independently by cell death assays and single cell transcriptomic analysis of MPP+ treated neural cultures. Together, these results highlight the importance of disease relevant cell type in stem cell modelling.
|
| |
|
| Overall design |
single cell RNA sequencing data of a time course differentiation of human induced Pluripotent Stem Cells (iPSC) into dopaminergic (DA) neurons (LMX1A Cre AAVS1 BFP line). The dataset consists of 8 samples, including 4 differentiation days (D21, D30, D45 and D65). For D21, D30 and D65, two different clones were sequenced (HET1/L25, HET2/L35). For D45, both samples are from the same clone (HET1/L25) but in one of the samples a nuclear isolation protocol was followed.
|
| |
|
| Contributor(s) |
Monzón-Sandoval J, F Cardo L, Li Z, Webber C, Li M |
| Citation(s) |
38132179 |
| |
| Submission date |
Nov 13, 2023 |
| Last update date |
Jan 02, 2024 |
| Contact name |
Jimena Monzón-Sandoval |
| E-mail(s) |
MonzonSandovalJ@cardiff.ac.uk
|
| Organization name |
UK Dementia Research Institute at Cardiff University
|
| Street address |
Maindy Road
|
| City |
Cardiff |
| ZIP/Postal code |
CF24 4HQ |
| Country |
United Kingdom |
| |
|
| Platforms (1) |
| GPL20301 |
Illumina HiSeq 4000 (Homo sapiens) |
|
| Samples (8)
|
|
| Relations |
| BioProject |
PRJNA1039802 |
Less...
More...