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Research Papers

High-Throughput Production of Single-Cell Microparticles Using an Inkjet Printing Technology

[+] Author and Article Information
Tao Xu, Helen Kincaid, Anthony Atala

Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157

James J. Yoo1

Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157jyoo@wfubmc.edu

1

Corresponding author.

J. Manuf. Sci. Eng 130(2), 021017 (Apr 11, 2008) (5 pages) doi:10.1115/1.2903064 History: Received December 14, 2007; Revised January 10, 2008; Published April 11, 2008

In this study, a novel biocompatible and inexpensive method for the rapid production of single-cell based microparticles is described. Using an HP DeskJet 550C printer, alginate microparticles containing one to several insulin-producing cells (beta-TC6) were fabricated by coprinting the cells and sodium alginate suspension into a CaCl2 solution. This method is able to generate microparticles of 3060μm in diameter at a rate as high as 55,000particless. Cell survival assays showed that more than 89% of printed cells survived the fabrication process. The printed beta-TC6 cells demonstrated continuous insulin secretion over a 6day period, which suggests that the printed cells are able to maintain normal cellular function within the microparticles. We show that the printing conditions, such as cell number, alginate concentration, and ionic strengths of CaCl2, influence cellular distribution and geometry of the printed particles. This study presents a simple and high-throughput method to encapsulate single cells, and this technique may be applied in various research investigations, including single-cell analysis, high-throughput drug screening, and stem cell differentiation at the single-cell level.

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Copyright © 2008 by American Society of Mechanical Engineers
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Figures

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Figure 1

Phase contrast images of printed beta-TC6 cell microparticles. (A) Many individual particles were dispersed in medium 1day after culture. Most particles contained single or just a few cells, and very few particles were without cells. (B) High magnification of a microparticle containing a single cell 1day after culture. (C) Individual microparticle contained beta-TC6 cells 1month after culture. The microparticle maintained its original structure, and the cells were still entrapped inside the particle. Magnifications are as follows: 100× (A) and 400× (B) and (C)

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Figure 2

The number of entrapped cells with respect to varying cell concentrations. As the cell concentration in the print solution increased, the number of printed particles containing cells increased (shown in gray bars). However, the number of microparticles containing only a single cell significantly decreased with increase in cell concentration (shown in black bars). (p*>0.01, n=10.)

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Figure 3

The effects of printing parameters on particle geometry. As CaCl2 ionic strength increased, the diameter of the printed particles increased (A). Increases in the alginate concentration did not significantly change the particle diameter (B). However, changes in the alginate concentration resulted in dramatic variation in the particle geometries (C). Most particles printed from 1% alginate showed round structures, while most particles generated from 0.5% alginate produced irregular shapes (C). Particles from 2% alginate showed long tailed structures (c). (p*>0.01, n=10). Magnification: 100×.

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Figure 4

Viability of the cell-loaded microparticles 1day after printing. The printed particles showed similar mean viability as the controls (p<0.05, n=10), which were prepared by manually seeding beta-TC6 cells onto standard tissue culture plates.

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Figure 5

The insulin secretion profiles for the cultures of beta-TC6 cell-containing microparticles and the controls over a 6day period. The printed beta-TC6 cells displayed a continuous insulin secretion during this time. The secreted insulin concentrations from the printed particles were comparable to that of the control group, which were prepared by manually seeding beta-TC6 cells onto standard tissue culture plates.

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