Publicação
Pluripotent stem cells as a tool to model Parkison’s disease in vitro
| Resumo: | Parkinson’s disease (PD) is the second most common neurodegenerative disease worldwide, typically characterized by the degeneration of the substantia nigra pars compacta (SNpc) and widespread pathology of Lewy-bodies. Multiple in vitro models have been used to address the phenotypical changes in the susceptible substantia nigra dopaminergic neurons and test potential neuroprotective drugs. Traditionally, changes have been triggered through genetic engineering approaches that induce the pathological effect of genetic mutations in PD or toxins that mimic environmentally induced phenotypes, usually in cell lines or primary cultures. However, the intrinsic flaws of these models are the low representation of dopaminergic neurons in culture, the technical challenges to genetically modify the genome, the high variability between primary cultures or the results reproducibility between the model original species and humans. Pluripotent stem cells specifically induced pluripotent stem cells have provided an alternative for developing models with high yield of dopaminergic neurons, both in 2D and 3D cultures, overcoming most of the drawbacks from other models. Here we explore pluripotent stem cells as a tool for the development of a dopaminergic neuron model by means of directed differentiation. For that we started by characterizing induced pluripotent stem cells for their stemness, then we directly differentiated pluripotent stem cells into dopaminergic neurons and established a 2D, 6-OHDA neurotoxin-based model for the screening of neuroprotective molecules. Moreover, we showed the biocompatibility of differentiated neurons with functionalized fluorescent nanodiamonds that may serve as a novel imaging tool for the dysfunction of dopaminergic neurons, and the growth of dopaminergic neurons in Ormocomp™ based structures that may serve as a grid for in vitro three-dimensional assessment of dopaminergic dysfunction, when combined with the functionalized nanodiamonds. |
|---|---|
| Autores principais: | Vedor, Carlos Eduardo Gonçalves |
| Assunto: | 6-OHDA Induced pluripotent stem cells Fluorescent nanodiamonds Parkinson’s disease Células estaminais pluripotentes induzidas Nanodiamantes fluorescentes Doença de Parkinson |
| Ano: | 2023 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso embargado |
| Instituição associada: | Universidade do Minho |
| Idioma: | inglês |
| Origem: | RepositóriUM - Universidade do Minho |
| Resumo: | Parkinson’s disease (PD) is the second most common neurodegenerative disease worldwide, typically characterized by the degeneration of the substantia nigra pars compacta (SNpc) and widespread pathology of Lewy-bodies. Multiple in vitro models have been used to address the phenotypical changes in the susceptible substantia nigra dopaminergic neurons and test potential neuroprotective drugs. Traditionally, changes have been triggered through genetic engineering approaches that induce the pathological effect of genetic mutations in PD or toxins that mimic environmentally induced phenotypes, usually in cell lines or primary cultures. However, the intrinsic flaws of these models are the low representation of dopaminergic neurons in culture, the technical challenges to genetically modify the genome, the high variability between primary cultures or the results reproducibility between the model original species and humans. Pluripotent stem cells specifically induced pluripotent stem cells have provided an alternative for developing models with high yield of dopaminergic neurons, both in 2D and 3D cultures, overcoming most of the drawbacks from other models. Here we explore pluripotent stem cells as a tool for the development of a dopaminergic neuron model by means of directed differentiation. For that we started by characterizing induced pluripotent stem cells for their stemness, then we directly differentiated pluripotent stem cells into dopaminergic neurons and established a 2D, 6-OHDA neurotoxin-based model for the screening of neuroprotective molecules. Moreover, we showed the biocompatibility of differentiated neurons with functionalized fluorescent nanodiamonds that may serve as a novel imaging tool for the dysfunction of dopaminergic neurons, and the growth of dopaminergic neurons in Ormocomp™ based structures that may serve as a grid for in vitro three-dimensional assessment of dopaminergic dysfunction, when combined with the functionalized nanodiamonds. |
|---|