Preliminary study on the construction of three-dimensional hippocampal neural network by using microfluidic technology <i>in vitro</i>_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

KONG Xianmin ¹ , TIAN Shanshan ² , CHEN Tao ² ,  HUANG Yinghui ¹

1. School of Biomedical Engineering 【？】, Beijing University of Technology, Beijing, 100124, P.R.China;
2. Institute of Laser Engineering, Beijing University of Technology, Beijing, 100124, P.R.China;

Corresponding author：

HUANG Yinghui, Email: yhuang@bjut.edu.cn

Keywords：

Microfluidic chip; neuron network; three-dimensional culture; hippocampal neuron

DOI：

10.7507/1002-1892.201809094

Video：

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Objective To preliminary study on the feasibility of constructing three-dimensional (3D) hippocampal neural network in vitro by using microfluidic technology.Methods A network patterned microfluidic chip was designed and fabricated by standard wet etching process. The primary hippocampal neurons of neonatal Sprague Dawley rats were isolated and cultured, and then inoculated on microfluidic chip for culture. Immunofluorescence staining was used to observe the growth of hippocampal neurons at 3, 5, and 7 days of culture and electrophysiological detection of hippocampal neuron network at 7 days of culture.Results The results showed that the number of hippocampal neurons increased gradually with the prolongation of culture time, and the neurite of neurons increased accordingly, and distributed uniformly and regularly in microfluidic chip channels, suggesting that the 3D hippocampal neuron network was successfully constructed in vitro. Single and multi-channel spontaneous firing signals of hippocampal neuronal networks could be detected at 7 days of culture, suggesting that neuronal networks had preliminary biological functions.Conclusion Patterned microfluidic chips can make hippocampal neurons grow along limited paths and form 3D neuron networks with corresponding biological functions such as signal transduction, which lays a foundation for further exploring the function of neuron networks in vitro.

Citation： KONG Xianmin, TIAN Shanshan, CHEN Tao, HUANG Yinghui. Preliminary study on the construction of three-dimensional hippocampal neural network by using microfluidic technology in vitro. Chinese Journal of Reparative and Reconstructive Surgery, 2019, 33(2): 239-242. doi: 10.7507/1002-1892.201809094 Copy

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1. Ng AH, Uddayasankar U, Wheeler AR. Immunoassays in microfluidic systems. Anal BioanalChem, 2010, 397(3): 991-1007.
2. Xu Y, Yang X, Wang E. Review:Aptamers in microfluidic chips. Anal ChimActa, 2010, 683(1): 12-20.
3. Varghese SS, Zhu Y, Davis TJ, et al. FRET for lab-on-a-chip devices-current trends and future prospects. Lab Chip, 2010, 10(11): 1355-1364.
4. Vincent ME, Liu W, Haney EB, et al. Microfiuidic stochastic confinement enhances analysis of rate cells by isolating cells and creating high density environments for control of diffusible signals. Chem Soc Rev, 2010, 39(3): 974-984.
5. Papp K, Végh P, Hóbor R, et al. Characterization of factors influencing on-chip complement activation to optimize parallel measurement of antibody and complement proteins on antigen microarrays. J Immunol Methods, 2011, 375(1-2): 75-83.
6. Toh A, Wang Z, Yang C, et al. Engineering microfluidic concentration gradient generators for biological applications. Microfluidics and Nanofluidics, 2014, 16(1): 1-18.
7. Plessy C, Desbois L, Fujii T, et al. Population transcriptomics with single-cell resolution: a new field made possible by microfluidics: a technology for high throughput transcript counting and data-driven definition of cell types. Bioessays, 2013, 35(2): 131-140.
8. Qin J, Ye N, Liu X, et al. Microfluidic devices for the analysis of apoptosis. Electrophoresis, 2005, 26(19): 3780-3788.
9. Sia SK, Whitesides GM. Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies. Electrophoresis, 2003, 24(21): 3563-3576.
10. Sanders G, Manz A. Chip-based microsystems for genomic and proteomic analysis. TrAC Trends in Analytical Chemistry, 2000, 19(6): 364-378.
11. ShenF, KastrupCJ, Ismagilov F. Using microfluidics to understand the effect of spatial distribution of tissue factor on blood coagulation. Thrombosis Research, 2008, 122(Suppl1): S27-S30.
12. Gomez N, Lu Y, Chen S, et al. Immobilized nerve growth factor and microtopography havedistinct effects on polarization versus axon elongation in hippocampal cells in culture. Biomaterials, 2007, 28(2): 271-284.
13. Taylor AM, Blurton-Jones M, Rhee SW, et al. A microfluidic culture platform for CNSaxonal injury, regenerationand transport. Nature Methods, 2005, 2(8): 599-605.
14. Christian KM, Song HJ, Ming GL. Functions and dysfunctions of adult hippocampalneurogenesis. Annual Review of Neuroscience, 2014, 37(1): 243.
15. Parihar VK, Limoli CL. Cranial irradiation compromises neuronal architecture in the hippocampus. Proc Natl Acad Sci USA, 2013, 110(31): 12822-12827.
16. Chakraborti A, Allen A, Allen B, et al. Cranial irradiation alters dendritic spine densityand morphology in the hippocampus. PLoS One, 2012, 7(7): e40844.

Chinese Journal of Reparative and Reconstructive Surgery

Preliminary study on the construction of three-dimensional hippocampal neural network by using microfluidic technology in vitro

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