Abstract
Various types of animal neurons were cultured on a microelectrode array (MEA) platform to form biosensors to detect potential environmental neurotoxins. For a large-scale screening tool, rodent MEA-based cortical-neuron biosensors would be very costly but chick forebrain neurons (FBNs) are abundant, cost-effective, and easy to dissect. However, chick FBNs have a lifespan of ~14 days in vitro and their spontaneous spike activity (SSA) has been difficult to develop and detect. We used a high-density neuron-glia co-culture on an MEA to prolong chick FBN lifetime to 3 months with lifetime-long SSA. A remarkable embryonic age-dependency in the culture’s morphology, lifespan, and most features of SSA signal was discovered. Our results show the feasibility of developing a chick FBN-MEA biosensor and also establish a new electrophysiological platform for functional study of an in vitro neuronal network.
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Abbreviations
- SSA:
-
Spontaneous spiking activity
- MEA:
-
Microelectrode array
- FBNs:
-
Forebrain neurons
- SCNs:
-
Spinal cord neurons
- M199:
-
Medium 199
- DIV:
-
Days in vitro
- En:
-
Embryonic day n
- ACC:
-
Active channel count, i.e., the number of active channels that are firing SSA signals from an MEA
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Acknowledgments
This work was partially supported by funding from the National Institutes of Health through SC COBRE (P20RR021949), the National Natural Science Foundation of China (Nos. 31070847 and 31370956), and the Strategic New Industry Development Special Foundation of Shenzhen (No. JCYJ20130402172114948). Serena Y. Kuang designed the study, did most of the forebrain tissue dissection, cell culturing, MEA recording, data processing tasks, and wrote the manuscript. Zhonghai Wang was responsible for MEA technical support and programmed the MatLab-based software (NeuroMEA) for MEA data processing. Ting Huang translated all MEA data processing requirements into a flowchart with programming logic that oriented the development of the NeuroMEA software. Lina Wei contributed to chick forebrain and spinal cord dissection and some MEA recordings. Mark Kindy served as a senior researcher in this study and modified the manuscript. As PIs of the Project, Drs. Tingfei Xi and Bruce Z. Gao supervised the overall experimental design and data interpretation and modified and finalized the manuscript.
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The authors declare no commercial or financial conflict of interest.
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Kuang, S.Y., Wang, Z., Huang, T. et al. Prolonging life in chick forebrain-neuron culture and acquiring spontaneous spiking activity on a microelectrode array. Biotechnol Lett 37, 499–509 (2015). https://doi.org/10.1007/s10529-014-1704-1
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DOI: https://doi.org/10.1007/s10529-014-1704-1