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

Authors

Alfredo Bellon, Department of Psychiatry and Behavioral Health, Penn State Hershey Medical Center, Hershey, PA 17033, USA.
Tuna Hasoglu, Department of Psychiatry and Behavioral Health, Penn State Hershey Medical Center, Hershey, PA 17033, USA.
Mallory Peterson, Department of Engineering Science and Mechanics, Penn State College of Engineering, State College, Philadelphia, PA 19107, USA.
Katherine Gao, Department of Psychiatry & Human Behavior, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Michael Chen MD, Lehigh Valley Health NetworkFollow
Elisabeta Blandin, Department of Neural & Behavioral Sciences, Penn State Hershey Medical Center, Hershey, PA 17033, USA.
Alonso Cortez-Resendiz, Department of Psychiatry and Behavioral Health, Penn State Hershey Medical Center, Hershey, PA 17033, USA.
Gary A. Clawson, Department of Pathology, Penn State University College of Medicine, Gittlen Cancer Research Laboratories, Hershey, PA 17033, USA.
Liyi Elliot Hong, Department of Psychiatry, Maryland Psychiatric Research Center, University of Maryland School of Medicine, Baltimore, MD 21201, USA.

Document Type

Article

Publication Date

10-20-2021

Publication Title

Brain sciences

ISSN

2076-3425

Department(s)

Department of Psychiatry; Fellows and Residents

Keywords

autism, biomarker, cytoskeleton, dendrite, neurite, neurodevelopment, schizophrenia, stem cells, transdifferentiation and neuronal model

Abstract

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.

Volume

11

Issue

11

DOI

10.3390/brainsci11111372

PubMed ID

34827371

Recommended Citation

Bellon, A., Hasoglu, T., Peterson, M., Gao, K., Chen, M., Blandin, E., Cortez-Resendiz, A., Clawson, G. A., & Hong, L. E. (2021). Optimization of Neurite Tracing and Further Characterization of Human Monocyte-Derived-Neuronal-like Cells. LVHN Scholarly Works. Retrieved from https://scholarlyworks.lvhn.org/research-historical-works/24
DOI: 10.3390/brainsci11111372