SURE 2014 Poster
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Transcript of SURE 2014 Poster
CELL PRINTER OPTIMIZATION FOR BIOMEMS APPLICATIONSMichael Lopez, Muhaimeen Hossain, Ashley Bui, Alec S.T. Smith, Catia Bernabini, Bonnie Berry, James J. Hickman
1NanoScience Technology Center, University of Central Florida, Orlando, FL, USA
Introduction
Methods
Results
References and
Acknowledgements
Future Plans
Conclusion
Drug development is an arduous process,
sometimes taking years to get a drug from research to the
market. Human clinical trials are expensive and often fail
despite positive outcomes obtained from animal models.
Moreover, the use of animals for drug testing carries
ethical concerns and coupled with difficulty in translating
results from animals to humans, highlights the need for
creating human-based platforms for reliable in vitro drug
testing. By creating a “body-on-a-chip” system, we can
efficiently test drugs on human systems while
simultaneously cutting the costs and the risks associated
with utilizing human subjects.
Currently, print parameters are being optimized for
use in printing a two-dimensional cell pattern on
BioMEMS (Biological Microelectromechanical Systems)
for electrophysiological testing. These tests will allow us
to monitor cell viability and revise our cell culturing
techniques. Combining various human cell types cultured
into one organized, in vitro system will produce a “human-
on-a-chip” product that has the capability to replace all
former methods of testing drug toxicity and physiological
response.
The reengineering and optimization of a cell printer
would pave the way for future experiments that seek to
change the way drugs are developed and tested.
Advantages of this printing include:
• Allowing the precise placement of cells on a
micrometer scale
• Capability to print multiple cell types within a single
device or substrate
• Ability to control the geometric placement of cells in
patterns determined by the user
Specific applications also include providing reliable
placement of muscle and neural cells for the formation of
neuromuscular junctions, allowing us to:
• Analyze the data retrieved from cells grown in vitro with
in vivo processes
• Follow the eventual path of printing of human cells
NanoScience Technology Center Hybrid Systems Laboratory
• RegenHU cell printer (Figure 2)
• C2C12 Mouse Myoblast Cell Lines
• Printed and maintained using DMEM with FBS (with
serum) or lab-developed serum free medium (M1)
• Once cells were confluent, differentiated using
DMEM with B27
• Testing of cell viability under various conditions:
• Humidity
• Temperature
• Cell Density
• Cell suspension settling time
• Print media such as serum, without serum,
hydrogels, PVP, and tissue culture mineral oil
Once conditions are optimized, printing will be done
on cantilevers to form neuromuscular junctions
accurate enough to be less than a millimeter apart.
Figure 3. C2C12 cultures printed in two types of media.
The above phase-contrast images are of printed C2C12
cells on glass cover slips coated with DETA. DMEM with
FBS (top) and with M1 (bottom) were the media used to
mature the cells. All of the cells were then differentiated in
DMEM with B27.
The ability to culture cells in vitro with comparable
properties to in vivo conditions is a progressive stride
within the medical field, specifically the pharmaceutical
industry. To maximize the effectiveness of drugs, it is
essential that proper testing models are available. Human
clinical trials are expensive and present numerous risks—
an in vitro “body-on-a-chip” model would significantly
reduce or eliminate the need for animal testing and would
significantly accelerate the drug development, testing, and
approval process.
Figure 4. Hygrometer display. Readout system of a
humidity sensor placed inside the sealed cell printer.
• The hygrometer has been programmed to measure
humidity once every two seconds, to ensure stability in
the system.
• Optimal humidity was determined to be 85 – 95%.
Figure 2. RegenHU Cell printer setup. The above setup of a cell printer (A) and computer (B) equipped with sophisticated
software capable of controlling various cell printing parameters. Specifically, the software is capable of controlling the X, Y, and
Z positions of the printhead while utilizing applications, such as BioCAD, to design print patterns.
Figure 5. Immunostained Neurons. The above confocal
images represent neurons stained for various segments of
their cell bodies. In the future, neurons will be printed at
optimal viability levels onto cantilevers and MEAs.
Figure 1. Cantilever Designs.
Neurons (red line) and Muscle
Cells (red dots) will be printed
onto cantilevers. This will provide
geometric placement of
appropriate cell types for the
formation of neuromuscular
junctions that are able to be
stimulated electronically.
RESULTS: Tests indicate optimal cell condition is printing C2C12s in serum containing medium
with a 5 minute cell adherence period in the printer under 85-95% humidity.
1. Huh, D. et al., (2012) Lab Chip. 12: 2156–2164
2. Song, H. et al., (2013) Lab Chip. 13: 1201
3. Smith, A. et al., (2014) Technology. 1: 37-48
This work was supported by NIH Grant Numbers R01 EB005459 and
UH2TR000516 .
GOAL: Optimize method of printing various cell
types onto chemically patterned substrates for
use in body-on-a-chip applications.