Block-Cell-Printing for live single-cell printing

Edited by James R. Heath, California Institute of Technology, Pasadena, CA, and accepted by the Editorial Board January 14, 2014 (received for review July 18, 2013)
February 10, 2014
111 (8) 2948-2953

Significance

The ability of printing single-cell arrays with high precision and efficiency, single-cell resolution, multiple cell types, and maintenance of cell viability and function is essential for cell function and heterogeneity measurement. It is still hard for current methods to completely satisfy the above requirements. We report a unique live-cell printing technique, Block-Cell-Printing, that allows for convenient, precise, multiplexed, and high-throughput printing of functional single-cell arrays. Block-Cell-Printing has a minimum turnaround time of 0.5 h, a maximum resolution of 5 µm, and close to 100% cell viability. This method has been applied to study cell communications in heterotypic cell pairs with controlled morphology, characterize cells’ abilities to extend their membranes, and print primary neurons.

Abstract

A unique live-cell printing technique, termed “Block-Cell-Printing” (BloC-Printing), allows for convenient, precise, multiplexed, and high-throughput printing of functional single-cell arrays. Adapted from woodblock printing techniques, the approach employs microfluidic arrays of hook-shaped traps to hold cells at designated positions and directly transfer the anchored cells onto various substrates. BloC-Printing has a minimum turnaround time of 0.5 h, a maximum resolution of 5 µm, close to 100% cell viability, the ability to handle multiple cell types, and efficiently construct protrusion-connected single-cell arrays. The approach enables the large-scale formation of heterotypic cell pairs with controlled morphology and allows for material transport through gap junction intercellular communication. When six types of breast cancer cells are allowed to extend membrane protrusions in the BloC-Printing device for 3 h, multiple biophysical characteristics of cells—including the protrusion percentage, extension rate, and cell length—are easily quantified and found to correlate well with their migration levels. In light of this discovery, BloC-Printing may serve as a rapid and high-throughput cell protrusion characterization tool to measure the invasion and migration capability of cancer cells. Furthermore, primary neurons are also compatible with BloC-Printing.

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Acknowledgments

We thank the Houston Methodist Research Institute SEM core facility for assistance with instrumentation and Dr. Dongfang Liu and Dr. Peilin Zheng (Baylor College of Medicine) for help in obtaining the high-resolution fluorescence images shown in SI Appendix, Fig. S19B. We also acknowledge funding support from the Cancer Prevention and Research Institute of Texas (CPRIT-R1007), NIH Grants NIH-CA180083 and NIH-DA035868, the Emily Herman Research Fund, the Department of Defense (W81XWH-11-02-0168), the Alliance of Nanohealth, and the Golfers Against Cancer Foundation.

Supporting Information

Appendix (PDF)
Supporting Information
Supporting Information (PDF)
Supporting Information
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Information & Authors

Information

Published in

The cover image for PNAS Vol.111; No.8
Proceedings of the National Academy of Sciences
Vol. 111 | No. 8
February 25, 2014
PubMed: 24516129

Classifications

Submission history

Published online: February 10, 2014
Published in issue: February 25, 2014

Keywords

  1. cell array
  2. cell communication
  3. protrusion profiling
  4. neuron patterning

Acknowledgments

We thank the Houston Methodist Research Institute SEM core facility for assistance with instrumentation and Dr. Dongfang Liu and Dr. Peilin Zheng (Baylor College of Medicine) for help in obtaining the high-resolution fluorescence images shown in SI Appendix, Fig. S19B. We also acknowledge funding support from the Cancer Prevention and Research Institute of Texas (CPRIT-R1007), NIH Grants NIH-CA180083 and NIH-DA035868, the Emily Herman Research Fund, the Department of Defense (W81XWH-11-02-0168), the Alliance of Nanohealth, and the Golfers Against Cancer Foundation.

Notes

This article is a PNAS Direct Submission. J.R.H. is a guest editor invited by the Editorial Board.

Authors

Affiliations

Kai Zhang
Department of Nanomedicine and
Department of Cell and Developmental Biology and
Chao-Kai Chou
Department of Molecular and Cellular Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, TX 77030; and
Xiaofeng Xia
Department of Systems Medicine and Bioengineering, Houston Methodist Research Institute, Houston, TX 77030;
Department of Radiology, Weill Medical College of Cornell University, New York, NY 10065;
Mien-Chie Hung
Department of Molecular and Cellular Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, TX 77030; and
Center for Molecular Medicine and Graduate Institute of Cancer Biology, China Medical University, Taichung 404, Taiwan
Department of Nanomedicine and
Department of Cell and Developmental Biology and
Department of Molecular and Cellular Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, TX 77030; and

Notes

1
To whom correspondence should be addressed. E-mail: [email protected].
Author contributions: K.Z. and L.Q. designed research; K.Z. performed research; K.Z., C.-K.C., X.X., M.-C.H., and L.Q. analyzed data; and K.Z. and L.Q. wrote the paper.

Competing Interests

The authors declare no conflict of interest.

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    Block-Cell-Printing for live single-cell printing
    Proceedings of the National Academy of Sciences
    • Vol. 111
    • No. 8
    • pp. 2857-3195

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