The
3D droplet bioprinter, developed by the Bayley Research Group at
Oxford, producing millimeter-sized tissues (credit: Sam Olof/ Alexander
Graham)
Scientists at the
University of Oxford
have developed a radical new method of 3D-printing laboratory-grown
cells that can form complex living tissues and cartilage to potentially
support, repair, or augment diseased and damaged areas of the body.
Printing high-resolution living tissues is currently difficult
because the cells often move within printed structures and can collapse
on themselves. So the team devised a new way to produce tissues in
protective nanoliter droplets wrapped in a lipid (oil-compatible)
coating that is assembled, layer-by-layer, into living cellular
structures.
3D-printing
cellular constructs. (left) Schematic of cell printing. The dispensing
nozzle ejects cell-containing bioink droplets into a lipid-containing
oil. The droplets are positioned by the programmed movement of the oil
container. The droplets cohere through the formation of droplet
interface lipid bilayers. (center) A related micrograph of a patterned
cell junction, containing two cell types, printed as successive layers
of 130-micrometer droplets ejected from two glass nozzles. (right) A
confocal fluorescence micrograph of about 700 printed human embryonic
kidney cells under oil at a density of 40 million cells per milliliter
(scale bar = 150 micrometers). (credit: Alexander D. Graham et
al./Scientific Reports)
This new method improves the survival rate of the individual cells
and allows for building each tissue one drop at a time to mimic the
behaviors and functions of the human body. The patterned cellular
constructs, once fully grown, can mimic or potentially enhance natural
tissues.
“We were aiming to fabricate three-dimensional living tissues that
could display the basic behaviors and physiology found in natural
organisms,” explained Alexander Graham, PhD, lead author and 3D
Bioprinting Scientist at OxSyBio (Oxford Synthetic Biology).*
“To date, there are limited examples of printed tissues [that] have
the complex cellular architecture of native tissues. Hence, we focused
on designing a high-resolution cell printing platform, from relatively
inexpensive components, that could be used to reproducibly produce
artificial tissues with appropriate complexity from a range of cells,
including stem cells.”
A
confocal micrograph of an artificial tissue containing two populations
of human embryonic kidney cells (HEK-293T) printed in the form of an
arborized structure within a cube (credit: Sam Olof/Alexander Graham)
The researchers hope that with further development, the materials
could have a wide impact on healthcare worldwide and bypass clinical
animal testing. The scientists plan to develop new complementary
printing techniques that allow for a wider range of living and hybrid
materials, producing tissues at industrial scale.
“We believe it will be possible to create personalized treatments by
using cells sourced from patients to mimic or enhance natural tissue
function,” said Sam Olof, PhD, Chief Technology Officer at OxSyBio. “In
the future, 3D bio-printed tissues may also be used for diagnostic
applications — for example, for drug or toxin screening.”
The study results were published August 1 in the open-access journal
Scientific Reports.
Abstract of High-Resolution Patterned Cellular Constructs by Droplet-Based 3D Printing
Bioprinting is an emerging technique for the fabrication of living
tissues that allows cells to be arranged in predetermined
three-dimensional (3D) architectures. However, to date, there are
limited examples of bioprinted constructs containing multiple cell types
patterned at high-resolution. Here we present a low-cost process that
employs 3D printing of aqueous droplets containing mammalian cells to
produce robust, patterned constructs in oil, which were reproducibly
transferred to culture medium. Human embryonic kidney (HEK) cells and
ovine mesenchymal stem cells (oMSCs) were printed at tissue-relevant
densities (10
7 cells mL
−1) and a high droplet
resolution of 1 nL. High-resolution 3D geometries were printed with
features of ≤200 μm; these included an arborised cell junction, a
diagonal-plane junction and an osteochondral interface. The printed
cells showed high viability (90% on average) and HEK cells within the
printed structures were shown to proliferate under culture conditions.
Significantly, a five-week tissue engineering study demonstrated that
printed oMSCs could be differentiated down the chondrogenic lineage to
generate cartilage-like structures containing type II collagen.
References:
- Alexander
D. Graham, Sam N. Olof, Madeline J. Burke, James P. K. Armstrong,
Ellina A. Mikhailova, James G. Nicholson, Stuart J. Box, Francis G.
Szele, Adam W. Perriman, Hagan Bayley. High-Resolution Patterned
Cellular Constructs by Droplet-Based 3D Printing. Scientific Reports,
2017; 7 (1) DOI: 10.1038/s41598-017-06358-x (open access)