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

被引：7

作者：

Bellon, Alfredo ^{[1
,2
]}

Hasoglu, Tuna ^{[1
]}

Peterson, Mallory ^{[3
]}

Gao, Katherine ^{[4
]}

Chen, Michael ^{[5
]}

Blandin, Elisabeta ^{[6
]}

Cortez-Resendiz, Alonso ^{[1
]}

Clawson, Gary A. ^{[7
]}

Hong, Liyi Elliot ^{[8
]}

机构：

[1] Penn State Hershey Med Ctr, Dept Psychiat & Behav Hlth, Hershey, PA 17033 USA

[2] Penn State Hershey Med Ctr, Dept Pharmacol, Hershey, PA 17033 USA

[3] Penn State Coll Engn, Dept Engn Sci & Mech, Philadelphia, PA 19107 USA

[4] Thomas Jefferson Univ, Dept Psychiat & Human Behav, Philadelphia, PA 19107 USA

[5] Lehigh Valley Hlth Network, Dept Psychiat, 2545 Schoenersville Rd, Bethlehem, PA 18017 USA

[6] Penn State Hershey Med Ctr, Dept Neural & Behav Sci, Hershey, PA 17033 USA

[7] Penn State Univ, Dept Pathol, Gittlen Canc Res Labs, Coll Med, Hershey, PA 17033 USA

[8] Univ Maryland, Sch Med, Maryland Psychiat Res Ctr, Dept Psychiat, Baltimore, MD 21201 USA

来源：

BRAIN SCIENCES | 2021年 / 11卷 / 11期

关键词：

schizophrenia; autism; stem cells; cytoskeleton; neurite; dendrite; neurodevelopment; biomarker; transdifferentiation and neuronal model; AUTISM SPECTRUM DISORDER; PLURIPOTENT STEM-CELLS; NEUROTRANSMITTER RELEASE; HIPPOCAMPAL-FORMATION; PREFRONTAL CORTEX; IN-VITRO; PROTEIN; SCHIZOPHRENIA; EXPRESSION; MAP2;

D O I：

10.3390/brainsci11111372

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

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.

引用

页数：16