고함량 스크리닝을 사용한 인간 유도만능줄기세포 유래 신경의 신경돌기 성장에 대한 신경독성 평가
Assessment of Neurotoxicity on Neurite Outgrowth in Human Induced Pluripotent Stem Cell-derived Neurons using High-content Screening
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Neurotoxicity refers to any adverse effect of exposure to a toxic chemical on the normal development of nervous system structure and/or functions, which has the potential to cause the disorder. The chemicals that need evaluation for neurotoxicity testing are increasing yearly. To screen for more efficient and high throughput methods for potential neurotoxicants, many researchers study the evaluation of a high-content screening system to detect chemical effects on neurite outgrowth in immortalized or tumor-derived neural cell lines of human origin or primary rodent neurons. However, these neural cells do not sufficiently represent the human nervous system. In this study, neuron was differentiated from human induced pluripotent stem cell (hiPSC), and by confirming neuronal function using the electrophysiological method. It was proved that the differentiated neurons are suitable as an alternative animal model. We identified the optimal conditions for measuring neurite outgrowth in hiPSC-derived neuron and evaluated two non-neurotoxicity chemicals and four neurotoxicity test chemicals. All four neurotoxicity chemicals decreased neurite outgrowth length, but two non-neurotoxicity chemicals not affected on neurite outgrowth length. Further, a set of chemicals (scopolamine hydrobromide and 2,5-hexanedione) were tested over a wide concentration range (0.1-100 μM). Scopolamine hydrobromide and 2,5-hexanedione not affected on neurite outgrowth length and cell viability. So far, we demonstrate that a high-content screening system can rapidly quantify chemical effects on neurite outgrowth in vitro. In addition, these studies will be useful for investigating the effects of other neurotoxic chemicals in vitro.
Abstract
Neurotoxicity refers to any adverse effect of exposure to a toxic chemical on the normal development of nervous system structure and/or functions, which has the potential to cause the disorder. The chemicals that need evaluation for neurotoxicity testing are increasing yearly. To screen for more efficient and high throughput methods for potential neurotoxicants, many researchers study the evaluation of a high-content screening system to detect chemical effects on neurite outgrowth in immortalized or tumor-derived neural cell lines of human origin or primary rodent neurons. However, these neural cells do not sufficiently represent the human nervous system. In this study, neuron was differentiated from human induced pluripotent stem cell (hiPSC), and by confirming neuronal function using the electrophysiological method. It was proved that the differentiated neurons are suitable as an alternative animal model. We identified the optimal conditions for measuring neurite outgrowth in hiPSC-derived neuron and evaluated two non-neurotoxicity chemicals and four neurotoxicity test chemicals. All four neurotoxicity chemicals decreased neurite outgrowth length, but two non-neurotoxicity chemicals not affected on neurite outgrowth length. Further, a set of chemicals (scopolamine hydrobromide and 2,5-hexanedione) were tested over a wide concentration range (0.1-100 μM). Scopolamine hydrobromide and 2,5-hexanedione not affected on neurite outgrowth length and cell viability. So far, we demonstrate that a high-content screening system can rapidly quantify chemical effects on neurite outgrowth in vitro. In addition, these studies will be useful for investigating the effects of other neurotoxic chemicals in vitro.
참고문헌 (27)
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