The Bio-Pick, Place and Perfuse (Bio-P3): A New Instrument for Building Organs
Faculty, Staff and Students
Jeff Morgan, Professor of Medical Science, BioMed
Anubhav Tripathi, Professor of Engineering, SOE
William Patterson III, Senior Research Engineer, SOE
Chris Bull, Director of Brown Design Workshop/Senior Lecturer, SOE
John Murphy III, Research Engineer, Technician, BioMed
Blanche Ip, Postdoctoral Research Associate, BioMed
Frank Cui, Biomedical Engineering Graduate Student, SOE
Kali Manning, Biomedical Engineering Graduate Student, BioMed
Benjamin Wilks, Biomedical Engineering Graduate Student, BioMed
Project Proposal: We are proposing an I-Team UTRA for three undergraduates to join our
interdisciplinary team of biologists, bioengineers, and engineers that are building a novel
instrument (Bio-Pick, Place & Perfuse, Bio-P3) that will pick up, place (stack) and fuse living
building parts to achieve layer-by-layer building of a thick living organ with high cell density. This
instrument is the next generation bio-printer and we are funded by a major instrument development
grant from the National Science Foundation.
Intellectual
Merit. The
major
engineering challenge to the field of
tissue engineering is the in vitro
fabrication of large solid organs with
high densities of living cells.
Diffusion of oxygen, nutrients and
removal of metabolic waste products
limit current engineered tissues to
thicknesses of ~100-200µm in order
to maintain cell viability. Natural
organs are much larger and contain a
branching vascular supply that
perfuses the entire organ and ensures Fig 1. Schematic of Bio P3 instrument. Step 1. Gripper head uses
all cells are close to blood vessels. As liquid suction to pick up a living part on the staging head. Step 2.
Stage moves build head into position under gripper. Step 3.
the field of tissue engineering Gripper deposits part on stack of living parts. Proto-organ is
struggles with this limitation, the perfused and environment monitored and controlled.
field of induced pluripotent (iPS)
stem cells is providing a plentiful source of immune-matched cells of a variety of tissues and
organs. We do not yet have a means for the in vitro fabrication of large 3D organs and tissues from
this source of cells. An instrument with this capability would have a worldwide impact in the field
of tissue engineering. Such an instrument would establish new paradigms in the fields of
biofabrication, biomanufacturing and would build new 3D models for research useful in any
number of basic as well as applied fields. The major challenges in this area of organ fabrication
are engineering in nature, albeit ones that must be informed by biology.
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Fundamentals of the Bio-P3. The Bio-P3 is a biological “pick and
place” device that will assemble/engineer large 3D tissues/organs
layer-by-layer using a controllable, low-level fluid suction head to
pick up living building parts and place them onto other building
parts in precise locations while maintaining perfusion as this living
structure is built. Our proof of principle instrument has been
published in the journal Tissue Engineering (1). This is a versatile
building platform that can grip multi-cellular building parts (of any
Fig 2. Honeycomb is a
size and shape), image the part it has gripped and then precisely
viable building unit.
place this part onto a stack of living building parts to effect the
Honeycomb (6 million
layer-by-layer engineering of a solid organ. Each living part has
cells) formed in microcarefully designed lumen structures and is composed of tens of
mold was stained live
(green)/dead (red) cells.
millions of cells formed in specific geometries designed to be
Staining shows viability.
stacked and used to build a large 3D tissue/organ complete with a
Bar 1800 μm. (1)
branched tubular (vascular) network for perfusion (Fig 1). Each
living part is designed to have lumens (holes) of different sizes and
when these building parts are stacked, their lumens are aligned to form a branching tubular
network that can be perfused. An example of such a building part that we have pioneered is a
large honeycomb structure from our paper in Biofabrication (Fig 2)(2). (First author is a
Brown undergraduate). She made this honeycomb-shaped building part by seeding monodispersed cells into specially designed non-adhesive agarose micro-molds. Within 24 hours, the
cells in this micro-mold aggregated and selfassembled a multi-cellular structure in the shape
of the honeycomb. The contiguous multicellular honeycomb formed around agarose
Fig 3. Toroid-shaped microtissues undergo fusion.
posts that directed the formation of lumens in
Bar 200μm.
the honeycomb. This tissue sheet is 2 cm endto-end in the x, y dimensions and less than 200µm in the z dimension. Thus, each cell in the
honeycomb receives adequate oxygen and nutrients because it is no more than 100µm away from
the top surface, bottom surface or the surface of a nearby lumen. Using computer aided design
and rapid prototyping, we design micro-molded hydrogels of virtually any size and shape and
these micro-molds direct self-assembly of living cells into the final shape of the building part.
We’ve shown that over 50 different cell types including primary human cells from a variety of
tissues and organs will form building parts. And further, we have shown in a Tissue Engineering
paper that these living building parts will readily fuse with one another to form even larger living
structures (Fig 3) (3) (First author is a Brown undergraduate who stayed for an ScM).
Shown are two toroid shaped parts of liver cells that have fused within 2 days.
Accomplishments:
 micro-molds to make living building parts of any design in 2 days
 living building parts that will self-fuse in 2 days to form larger structures
 version 1.0 Bio-P3 manual instrument that has stacked & perfused 20 honeycombs
 NSF funding to build new automated instrument
 assembled interdisciplinary team of faculty, staff and students
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Work underway:
 version 2.0 Bio-P3 under construction with automation and video monitoring
 joystick control of gripper head (x, y, z, rotation around z) pick & place of parts
 design, fabrication and characterization of gripper head, build head, build box
 environmental control and sensing: temperature, CO2, pH, O2
 design and characterization of new living building parts with capillary networks
Proposed work for Bio-P3 I-Team. We are seeking a diverse group of three 1st or 2nd year
undergraduates to join our interdisciplinary team. No experience necessary except desired
interests/affinities in one or more of the following: biology, medicine, bioengineering,
mechanical engineering, electrical engineering, computer science, mathematical
simulations, and economics.
Through faculty/student discussions, assigned readings, review of the scientific literature and
student presentations, the entire Bio-P3 I-TEAM will learn about:
 the growing shortage of organs needed for transplantation
 the medical, societal and economic impact of this shortage
 the organ donation process and the organ procurement/banking system
 the ethics of organ donation and the black market in organs
Through faculty/student/staff instruction, hands-on learning, supplemented with training videos,
Bio-P3 I-TEAM members, depending on their interests/affinities, will acquire several of the
following skills:
 Solidworks software design and printing of 3D prototypes
 Labview software control over devices including pumps, controllers, sensors
 COMSOL simulations of fluid dynamics
 ImageJ software for quantitative image analysis
 Culture of mammalian cells, cell counting, cell passage, cell cryopreservation
 Formation of 3D living building parts
 Brightfield, phase contrast, fluorescent and time lapse microscopy
 Biochemical analyses of living building parts and the performance of bioengineered
proto-organs
Our learning objectives for the Bio-P3 I-TEAM are to:
1. Understand the context of the societal need to find solutions to the shortage of organs.
2. Become proficient in valuable lab and engineering skills.
3. Apply these skills to the interdisciplinary effort to build a revolutionary instrument.
References
1. Blakely, A.M., Manning, K.L., Tripathi, A. and Morgan, J.R. Bio-Pick, Place and Perfuse: A
New Instrument for 3D Tissue Engineering. Tissue Eng. 21: 737-746, 2015.
2. Tejavibulya, N., Youssef, J., Bao, B., Ferruccio, T-M and Morgan, J.R. Directed SelfAssembly of Large Scaffold-free Multi-cellular Honeycomb Structures. Biofabrication 3, 1-9,
2011.
3. Livoti, C.M., and Morgan, J.R. Self-Assembly and Tissue Fusion of Toroid-Shaped Minimal
Building Units. Tissue Eng. 16: 2051-2061, 2010.
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