Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells

Deficits in neuronal structure are consistently associated with neurodevelopmental illnesses such as autism and schizophrenia. Nonetheless, the inability to access neurons from clinical patients has limited the study of early neurostructural changes directly in patients’ cells. This obstacle has bee...

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Autores principales:	Alfredo Bellon, Tuna Hasoglu, Mallory Peterson, Katherine Gao, Michael Chen, Elisabeta Blandin, Alonso Cortez-Resendiz, Gary A. Clawson, Liyi Elliot Hong
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	schizophrenia autism stem cells cytoskeleton neurite dendrite Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
Acceso en línea:	https://doaj.org/article/33c5f5cd5db8419ea8c833663d2e5ed4
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spelling	oai:doaj.org-article:33c5f5cd5db8419ea8c833663d2e5ed42021-11-25T16:55:59ZOptimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells10.3390/brainsci111113722076-3425https://doaj.org/article/33c5f5cd5db8419ea8c833663d2e5ed42021-10-01T00:00:00Zhttps://www.mdpi.com/2076-3425/11/11/1372https://doaj.org/toc/2076-3425Deficits in neuronal structure are consistently associated with neurodevelopmental illnesses such as autism and schizophrenia. Nonetheless, the inability to access neurons from clinical patients has limited the study of early neurostructural changes directly in patients’ cells. This obstacle has been circumvented by differentiating stem cells into neurons, although the most used methodologies are time consuming. Therefore, we recently developed a relatively rapid (~20 days) protocol for transdifferentiating human circulating monocytes into neuronal-like cells. These monocyte-derived-neuronal-like cells (MDNCs) express several genes and proteins considered neuronal markers, such as MAP-2 and PSD-95. In addition, these cells conduct electrical activity. We have also previously shown that the structure of MDNCs is comparable with that of human developing neurons (HDNs) after 5 days in culture. Moreover, the neurostructure of MDNCs responds similarly to that of HDNs when exposed to colchicine and dopamine. In this manuscript, we expanded our characterization of MDNCs to include the expression of 12 neuronal genes, including tau. Following, we compared three different tracing approaches (two semi-automated and one automated) that enable tracing using photographs of live cells. This comparison is imperative for determining which neurite tracing method is more efficient in extracting neurostructural data from MDNCs and thus allowing researchers to take advantage of the faster yield provided by these neuronal-like cells. Surprisingly, it was one of the semi-automated methods that was the fastest, consisting of tracing only the longest primary and the longest secondary neurite. This tracing technique also detected more structural deficits. The only automated method tested, Volocity, detected MDNCs but failed to trace the entire neuritic length. Other advantages and disadvantages of the three tracing approaches are also presented and discussed.Alfredo BellonTuna HasogluMallory PetersonKatherine GaoMichael ChenElisabeta BlandinAlonso Cortez-ResendizGary A. ClawsonLiyi Elliot HongMDPI AGarticleschizophreniaautismstem cellscytoskeletonneuritedendriteNeurosciences. Biological psychiatry. NeuropsychiatryRC321-571ENBrain Sciences, Vol 11, Iss 1372, p 1372 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	schizophrenia autism stem cells cytoskeleton neurite dendrite Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571
spellingShingle	schizophrenia autism stem cells cytoskeleton neurite dendrite Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Alfredo Bellon Tuna Hasoglu Mallory Peterson Katherine Gao Michael Chen Elisabeta Blandin Alonso Cortez-Resendiz Gary A. Clawson Liyi Elliot Hong Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells
description	Deficits in neuronal structure are consistently associated with neurodevelopmental illnesses such as autism and schizophrenia. Nonetheless, the inability to access neurons from clinical patients has limited the study of early neurostructural changes directly in patients’ cells. This obstacle has been circumvented by differentiating stem cells into neurons, although the most used methodologies are time consuming. Therefore, we recently developed a relatively rapid (~20 days) protocol for transdifferentiating human circulating monocytes into neuronal-like cells. These monocyte-derived-neuronal-like cells (MDNCs) express several genes and proteins considered neuronal markers, such as MAP-2 and PSD-95. In addition, these cells conduct electrical activity. We have also previously shown that the structure of MDNCs is comparable with that of human developing neurons (HDNs) after 5 days in culture. Moreover, the neurostructure of MDNCs responds similarly to that of HDNs when exposed to colchicine and dopamine. In this manuscript, we expanded our characterization of MDNCs to include the expression of 12 neuronal genes, including tau. Following, we compared three different tracing approaches (two semi-automated and one automated) that enable tracing using photographs of live cells. This comparison is imperative for determining which neurite tracing method is more efficient in extracting neurostructural data from MDNCs and thus allowing researchers to take advantage of the faster yield provided by these neuronal-like cells. Surprisingly, it was one of the semi-automated methods that was the fastest, consisting of tracing only the longest primary and the longest secondary neurite. This tracing technique also detected more structural deficits. The only automated method tested, Volocity, detected MDNCs but failed to trace the entire neuritic length. Other advantages and disadvantages of the three tracing approaches are also presented and discussed.
format	article
author	Alfredo Bellon Tuna Hasoglu Mallory Peterson Katherine Gao Michael Chen Elisabeta Blandin Alonso Cortez-Resendiz Gary A. Clawson Liyi Elliot Hong
author_facet	Alfredo Bellon Tuna Hasoglu Mallory Peterson Katherine Gao Michael Chen Elisabeta Blandin Alonso Cortez-Resendiz Gary A. Clawson Liyi Elliot Hong
author_sort	Alfredo Bellon
title	Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells
title_short	Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells
title_full	Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells
title_fullStr	Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells
title_full_unstemmed	Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells
title_sort	optimization of neurite tracing and further characterization of human monocyte-derived-neuronal-like cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/33c5f5cd5db8419ea8c833663d2e5ed4
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Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells

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