Bio-Pick, Place, and Perfuse: A New Instrument for Three-Dimensional Tissue Engineering

Abstract

A grand challenge of tissue engineering is the fabrication of large constructs with a high density of living cells. By adapting the principles of pick-and-place machines used in the high-speed assembly of electronics, we have developed an innovative instrument, the Bio-Pick, Place, and Perfuse (Bio-P3), which picks up large complex multicellular building parts, transports them to a build area, and precisely places the parts at desired locations while perfusing the parts. These assembled parts subsequently fuse to form a larger contiguous tissue construct. Multicellular microtissues were formed by seeding cells into nonadhesive micro-molds, wherein cells self-assembled scaffold-free parts in the shape of spheroids, toroids, and honeycombs. After removal from the molds, the parts were gripped, transported (using an x, y, z controller), and released using the Bio-P3 with little to no effect on cell viability or part structure. As many as 16 toroids were stacked over a 170 μm diameter post where they fused over the course of 48 h to form a single tissue. Larger honeycomb parts were also gripped and stacked onto a build head that, like the gripper head, provided fluid suction to hold and perfuse the parts during assembly. Scaffold-free building parts help to address several of the engineering and biological challenges to large tissue biofabrication, and the Bio-P3 described in this article is a novel instrument for the controlled gripping, placing, stacking, and perfusing of living building parts for solid organ fabrication.

Principles of the Kenzan Method for Robotic Cell Spheroid-Based Three-Dimensional Bioprinting

“…Based on the discussion above, this was a good option to signal that there is a difference in methodology from the regular scaffold-based 3D bioprinting. From a commercial standpoint, this designation also makes sense due to the fact that the target user groups are largely the same as with the bioprinting, and because other bio-assembling methods, like the 'bio-pick, place and perfuse' instrument 19 , exist although are less known.…”

Laser-Etched Designs for Molding Hydrogel-Based Engineered Tissues

“…Engineered tissues formed in these molds may be used as tissue units for a self-assembled building block approach, as proposed by Gwyther et al 44 Alternatively, the engineered tissue building blocks may be assembled into larger structures with controlled orientation by use of innovative technologies, such as the Bio-Pick, Place, and Perfuse (Bio-P3) instrument. 45 Finally, the laser-etched molds may be designed with 3D features able to integrate and confine polymeric fibers, meshes, or sponges to reinforce the constructs and further modulate their internal structure and porosity. [46][47][48]…”

Computational fluid dynamic analysis of bioprinted self‐supporting perfused tissue models

“…The scaffold‐dependent tissue is therefore dependent on these scaffolds to hold the tissue together over the long‐term. On the other hand, scaffold‐free is normally used to describe tissues that are comprised of cells embedded within their own cell‐secreted matrix (not contrived or reconstituted ECM scaffolds) and are therefore “scaffold‐free” (Aguilar, Olivos et al, ; Aguilar, Smith et al, ; Blakely, Manning, Tripathi, & Morgan, ; Itoh et al, ; Moldovan, ; Moldovan et al, ; Murata et al, ; Ozbolat, ; Toratani et al, ). The distinction between scaffold‐dependent and scaffold‐free can similarly be defined by the tissue's final composition after maturation.…”