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Transplantation of GABAergic precursor neurons generated from hESCs in postnatal day 2 mice

Hatamian, Diana (2020) MOBN03 20192
Degree Projects in Molecular Biology
Abstract
The World Health Organization has reported around 50 million people worldwide suffer from epilepsy. Epilepsy is a neurological disorder caused by abnormal brain activity that leads to spontaneous seizures. The complications that arise with epilepsy can heavily limit the quality of life for those affected and can lead to death. The seizures that occur are due to an imbalance of hyper excitatory neuronal activity and a lack of inhibitory neuronal response. There have been beneficial advancements in cell therapies that have great promise in treating epilepsy compared to previous therapeutics. Using GABAergic precursors derived from human embryonic stem cells (hESCs) made in vitro, we performed a pilot study transplanting these cells into the... (More)
The World Health Organization has reported around 50 million people worldwide suffer from epilepsy. Epilepsy is a neurological disorder caused by abnormal brain activity that leads to spontaneous seizures. The complications that arise with epilepsy can heavily limit the quality of life for those affected and can lead to death. The seizures that occur are due to an imbalance of hyper excitatory neuronal activity and a lack of inhibitory neuronal response. There have been beneficial advancements in cell therapies that have great promise in treating epilepsy compared to previous therapeutics. Using GABAergic precursors derived from human embryonic stem cells (hESCs) made in vitro, we performed a pilot study transplanting these cells into the hippocampus of postnatal 2-day old mice. After determining pup hippocampal coordinates, differentiating the GABAergic precursor neurons, and transplanting the cells in the hippocampus of the neonatal mice, we saw cell survival 2-months after transplantation. This pilot study is crucial for future development of cell transplantations that target the hippocampus of neonatal mice. (Less)
Popular Abstract
The World Health Organization (WHO) has recorded that around 50 million people worldwide suffer from epilepsy. It has been shown to be third the largest neurological disorder that affects people of all ages in terms of burden of disease. Epilepsy is a disorder in which brain activity becomes abnormal, causing seizures or periods of unusual behavior, sensations, and sometimes loss of awareness. So far, the best therapeutic options have been anti-epileptic drugs (AEDs) and brain tissue removal, but both have life-threatening side effects, and both only treat the neurological aspects of the disorder, not the neuropsychiatric aspects. Developments in stem cell therapy on the other hand have had preliminary experimental success in treating both... (More)
The World Health Organization (WHO) has recorded that around 50 million people worldwide suffer from epilepsy. It has been shown to be third the largest neurological disorder that affects people of all ages in terms of burden of disease. Epilepsy is a disorder in which brain activity becomes abnormal, causing seizures or periods of unusual behavior, sensations, and sometimes loss of awareness. So far, the best therapeutic options have been anti-epileptic drugs (AEDs) and brain tissue removal, but both have life-threatening side effects, and both only treat the neurological aspects of the disorder, not the neuropsychiatric aspects. Developments in stem cell therapy on the other hand have had preliminary experimental success in treating both aspects of the disorder.

Epileptic seizures occur when there is an imbalance of inhibition and excitation in the brain, specifically hyper excitation and not enough inhibition. This can occur dur to an interference with the function of sodium channels, which create action potentials, or blocking of calcium channels, which reduce neurotransmission. The aim of this study was to, first, differentiate human embryonic stem cells (hECSs) to GABAergic neurons (inhibitory neurons). We performed a cell differentiation using 2 transcription factors, Ascl1 and Dlx2, that have been shown to create GABAergic neurons. These transcription factors were introduced to the hESCs via lentiviral particles carrying doxycycline-inducible systems for Ascl1 and Dlx2. At 7 days in-vitro (DIV) and 14 DIV we collected the GABAergic precursor cells derived from the hESCs and transplanted them in the hippocampi of post-natal 2 day pups.

After the time span of 2-weeks and 2-months post transplantation we performed immunohistochemistry on the mouse brain tissue in order to stain for human neuronal cell markers and immune rejection markers to determine if the transplantation was successful. We were able to see transplanted cells at both time points, and evidence in the staining’s to prove the transplanted cells were not rejected by the host's immune system. Not only were the cells in the hippocampi where they had been transplanted, they had also migrated to other areas of the brain. Next steps could be deriving specific GABAergic interneurons from hESCs and transplanting those precursors into an epileptic model. In conclusion, the further development of cell replacement therapy as a treatment for neurological disorders such as epilepsy is a very promising field.


Master’s Degree Project in Molecular Biology, Molecular Genetics and Biotechnology, 60 credits
Department of Biology, Lund University

Advisor: My Andersson
Cellular Neurophysiology and Epilepsy, Epilepsy Center, BMC, Lund University (Less)
Please use this url to cite or link to this publication:
author
Hatamian, Diana
supervisor
organization
course
MOBN03 20192
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
9028451
date added to LUP
2020-09-07 14:25:00
date last changed
2020-09-07 14:25:00
@misc{9028451,
  abstract     = {{The World Health Organization has reported around 50 million people worldwide suffer from epilepsy. Epilepsy is a neurological disorder caused by abnormal brain activity that leads to spontaneous seizures. The complications that arise with epilepsy can heavily limit the quality of life for those affected and can lead to death. The seizures that occur are due to an imbalance of hyper excitatory neuronal activity and a lack of inhibitory neuronal response. There have been beneficial advancements in cell therapies that have great promise in treating epilepsy compared to previous therapeutics. Using GABAergic precursors derived from human embryonic stem cells (hESCs) made in vitro, we performed a pilot study transplanting these cells into the hippocampus of postnatal 2-day old mice. After determining pup hippocampal coordinates, differentiating the GABAergic precursor neurons, and transplanting the cells in the hippocampus of the neonatal mice, we saw cell survival 2-months after transplantation. This pilot study is crucial for future development of cell transplantations that target the hippocampus of neonatal mice.}},
  author       = {{Hatamian, Diana}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Transplantation of GABAergic precursor neurons generated from hESCs in postnatal day 2 mice}},
  year         = {{2020}},
}