Establishing a protocol for in vitro generation of GABAergic interneurons from human iPSCs
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- Tez No: 839094
- Danışmanlar: PROF. DR. MAGDALENA GÖTZ, PROF. DR. WOLFGANG WURST
- Tez Türü: Yüksek Lisans
- Konular: Tıbbi Biyoloji, Biyomühendislik, Medical Biology, Bioengineering
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2023
- Dil: İngilizce
- Üniversite: Technısche Unıversıtat Munchen
- Enstitü: Yurtdışı Enstitü
- Ana Bilim Dalı: Belirtilmemiş.
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: Belirtilmemiş.
Özet
GABAergic interneurons regulate cortical neural networks by providing inhibitory inputs. Their loss or dysfunction in the cerebral cortex leads to neurodevelopmental disorders such as schizophrenia, autism spectrum disorders, and epilepsy. Therefore, there has been a growing interest in the reproducible and highly efficient generation of cortical GABAergic interneurons. In vitro generation of GABAergic interneurons from human pluripotent stem cells (hPSCs) is crucial for studying human-specific cellular and functional properties during development, which cannot be modeled in other species with lower complexity. Also, these in vitro derived cells are good candidates for future therapeutic cell replacement therapies. Here I describe a protocol for generating cortical interneurons from hiPSCs towards medial ganglionic eminence (MGE)-derived ventral progenitors and further into GABAergic interneurons. I have established the protocol by comparing and combining several published protocols with the goal of achieving high percentage of GABAergic interneurons which can also be scaled up to obtain a large number of cells. I verified the identity of cells via staining and RT-qPCR at different time points (progenitor and neuronal stages) for a panel of medial ganglionic eminence (MGE), GABAergic, and neuronal markers. These showed a purity of up to 80% of my differentiation protocol. I further explored the functionality of the generated neurons running migration assays in vitro and ex vivo to observe their distinct migration behavior, as they migrate tangentially in the brain, also in comparison to the cortical projection neurons, which migrate radially in the cortex migration. Overall, I have established a reproducible protocol which generates 80% GABAergic markers able to migrate in vitro.
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