Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)

Authors

Ze-Zhi Wu, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Zheng-Wei Wang, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Li-Guang Zhang, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Zhi-Xing An, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Dong-Huo Zhong, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Qi-Ping Huang, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Mei-Rong Luo, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Yan-Jian Liao, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Liang Jin, Key Laboratory of Biorheological Science and Technology of the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, People’s Republic of China
Chen-Zhong Li, Florida International University, Nanobioengineering/Bioelectronics Laboratory, Department of Biomedical EngineeringFollow
William S. Kisaalita, University of Georgia, Cellular Bioengineering Laboratory, College of Engineering

Date of this Version

1-3-2013

Document Type

Article

Abstract

Introduction:

In this study, quasi-three-dimensional (3D) microwell patterns were fabricated with poly (l-lactic acid) for the development of cell-based assays, targeting voltage-gated calcium channels (VGCCs).

Methods and materials:

SH-SY5Y human neuroblastoma cells were interfaced with the microwell patterns and found to grow as two dimensional (2D), 3D, and near two dimensional (N2D), categorized on the basis of the cells’ location in the pattern. The capability of the microwell patterns to support 3D cell growth was evaluated in terms of the percentage of the cells in each growth category. Cell spreading was analyzed in terms of projection areas under light microscopy. SH-SY5Y cells’ VGCC responsiveness was evaluated with confocal microscopy and a calcium fluorescent indicator, Calcium Green^TM-1. The expression of L-type calcium channels was evaluated using immunofluorescence staining with DM-BODIPY.

Results:

It was found that cells within the microwells, either N2D or 3D, showed more rounded shapes and less projection areas than 2D cells on flat poly (l-lactic acid) substrates. Also, cells in microwells showed a significantly lower VGCC responsiveness than cells on flat substrates, in terms of both response magnitudes and percentages of responsive cells, upon depolarization with 50 mM K⁺. This lower VGCC responsiveness could not be explained by the difference in L-type calcium channel expression. For the two patterns addressed in this study, N2D cells consistently exhibited an intermediate value of either projection areas or VGCC responsiveness between those for 2D and 3D cells, suggesting a correlative relation between cell morphology and VGCC responsiveness.

Conclusion:

These results suggest that the pattern structure and therefore the cell growth characteristics were critical factors in determining cell VGCC responsiveness and thus provide an approach for engineering cell functionality in cell-based assay systems and tissue engineering scaffolds.

Comments

This article was originally published in International Journal of Nanomedicine.

Creative Commons License

This work is licensed under a Creative Commons Attribution 3.0 License.

Recommended Citation

Wu, Ze-Zhi; Wang, Zheng-Wei; Zhang, Li-Guang; An, Zhi-Xing; Zhong, Dong-Huo; Huang, Qi-Ping; Luo, Mei-Rong; Liao, Yan-Jian; Jin, Liang; Li, Chen-Zhong; and Kisaalita, William S., "Responsiveness of voltage-gated calcium channels in SH-SY5Y human neuroblastoma cells on quasi-three-dimensional micropatterns formed with poly (l-lactic acid)" (2013). Department of Biomedical Engineering Faculty Publications. 5.
https://digitalcommons.fiu.edu/biomed_eng/5