Materials and liquid handling system for cultivated and plant-based foods

Abstract

The disclosure relates to materials and liquid-handling system of print heads for use in the bioprinting of biostructures having predetermined two-dimensional (2D) and / or three- dimensional (3D) pattern of edible structures. Specifically, the disclosure is directed to an materials and liquid-handling system of print heads operable for the fabrication of edible biostructures using drop-on-demand dispensers.

Description

MATERIALS AND LIQUID HANDLING SYSTEM FOR CULTIVATED AND PLANT-BASED FOODSBACKGROUND

[0001] The disclosure is directed to materials and liquid-handling system of print heads for use in the bioprinting of biostructures having predetermined two-dimensional (2D) and / or three- dimensional (3D) pattern of edible structures. Specifically, the disclosure is directed to an materials and liquid-handling system of print heads operable for the fabrication of edible biostructures using drop-on-demand dispensers.

[0002] Three-dimensional (3D) bioprinting refers to a printing process in which patterns of biological materials whereby cells, or plant based proteins, in combination with, for example, hydrocolloids are deposited using three-dimensional printing technologies in such a way that functions of the bioprinted materials are developed to emulate a biological organ or tissue, and imitate biological fibers by plant-based proteins. Some of the technologies that are used for 3D bioprinting of cells are photolithography, magnetic bioprinting, stereolithography, and direct cell extrusion. Once printed, the bioprinted pre-tissue is transferred to an incubator and the cell-based pre-tissue matures into a tissue. The biological material (e.g. cells), however, can be suspended within a hydrogel, which helps to support and / or promote cell differentiation, assembly and tissue growth. A hydrogel is a liquid that becomes a gelatinous solid when exposed to a curing agent, or a change in its environment, such as temperature and / or pH. Curing agents can include UV light, cross-linking polymers, or chemicals such as calcium chloride.

[0003] It is difficult to print biological materials with hydrogels with specificity and precision due to the mixing effects between the curing agent and the biological materials. In addition, the timing of the mixing poses a substantial problem in standard bioprinting, as curing effects are time dependent as well, meaning the mixing profile becomes a limiting factor, often with few potential optimization points. In addition, the printing process itself can be inefficient. An additional challenge remains in the ability to get patterned printout of the cells, and the ability to precisely dispense the curing agent(s) or additional bio-inks.

[0004] Therefore, the need exists for materials and liquid-handling system with print heads (or dispensers) , dedicated to the use in the bioprinting of precision-fabricated bio structures, andcultured foods with plant-based components such as skeletal muscles to be used as edible protein sources capable of emulating recognizable complex tissue cuts of meat.SUMMARY

[0005] Disclosed, in various implementations, are assemblies of print heads for use in the bioprinting of biostructures having predetermined two (2D)- and / or three dimensional (3D) pattern of cells. Specifically, disclosed herein, are assemblies of print heads operable in bioprinting systems for the fabrication of edible biostructures using fully digital drop-on-demand dispensers.

[0006] In an exemplary implementation provided herein is a computerized printing station for a bioprinter comprising: a shelf sub-assembly, a back plate defining a longitudinal axis; coupled to the shelf sub-assembly, having a powertrain operable to vertically translate the shelf sub-assembly; a plurality of bio-ink buffer tanks coupled to the shelf sub assembly, each of the plurality of bio-ink buffer tanks operably coupled to a group of print heads, wherein each of the plurality of bio-ink buffer tanks is in communication with an adjacent buffer tank, and wherein a first buffer tank is in further communication with a first bio-ink reservoir; a frame supporting an array of a plurality of print heads divided into the groups, wherein the frame defines a longitudinal axis and is coupled to the shelf subassembly; a plurality of pressurized gas manifold pairs, coupled to a corresponding group, wherein each of the plurality of pressurized gas manifolds are each being in communication with a corresponding half pair of the pressurized gas manifold of an adjacent group, and wherein a first pair of gas manifolds is in further liquid communication with a first pressurized gas reservoir; a conveyor, operable to convey a substrate in parallel with the longitudinal axis defined by the back plate; and a central processing module (CPM), in communication with the back plate powertrain, the plurality of bio-ink buffer tanks, the first compressed gas reservoir, and each of the plurality of print heads, the CPM further comprising at least one processor in communication with a non-transitory memory device storing thereon a computer-readable medium with a set of executable instructions configured, when executed by the at least one processor to operate the printing station.

[0007] In another exemplary implementation, provided herein is a bioprinter comprising: a plurality of the printing stations disclosed herein; and a central command and control module operable to control the plurality of printing stationsBRIEF DESCRIPTION OF THE FIGURES

[0008] For a better understanding of the print heads assembly operable in a bioprinting systems for the fabrication of edible non-random biostructures using drop-on-demand of at least one bio-ink, with regard to the exemplary implementations thereof, reference is made to the accompanying examples and figures, in which:

[0009] FIG.1A, illustrates the front top perspective view of an exemplary implementation of print head assembly, with FIG. IB illustrating a bottom perspective thereof, while FIG. 1C, illustrating a side perspective thereof, and FIG. ID illustrating an enlarged portion of the bottom perspective view in FIG. IB;

[0010] FIG. 2A, illustrates the front top perspective of an exemplary implementation of a single print head, with FIG. 2B, illustrating a bottom rear perspective thereof, with FIG. 2C, illustrating nozzle offset of the dispensers as viewed from direction A illustrated in FIG. 2B;

[0011] FIG. 3, illustrates a X-Z cross section of the intermediate bio-ink buffer tank;

[0012] FIG. 4, is a schematic showing the configuration of extra flight distances for an unlevelled build plate; and

[0013] FIG. 5A, is a schematic showing degrees of freedom between the print tray and a single print head band, with FIG. 5B, being a schematic showing degrees of freedom between the print tray and an array of print heads.DETAILED DESCRIPTION

[0014] Provided herein are exemplary implementations of materials and liquid-handling system of print heads for use in the bioprinting of cultivated food components printing having predetermined two (2D) - and / or three-dimensional (3D) patterns, to emulate the structure and fibers of edible animal products. Specifically, provided herein are exemplary implementations of assemblies of print heads in a single framework operable in a bioprinting systems for the fabrication of edible biostructures using drop-on-demand dispensers.Definition.

[0015] In the context of the disclosure, the term “bioprinting” refers to a process depositing material mixed with cells and / or other bio-inks using for example, inkjet printer having drop-on- demand capabilities, fused filament deposition, extrusion or molding. Accordingly, the term “bioprinting” means the deposition of predetermined compositions for the purpose of forming, oremulating any structure / tissue / organ, which is composed of more than one cell, present in mammalian body, having some kind of organization and / or complexity and / or functionality. A mere random cluster of cells therefore would not fall under this definition. However, a composition, which has no mammalian, cells, but is rather plant-based proteins that do emulate structure / tissue / organ present in mammalian, piscine, or avian body- does fall under this definition. In this context, “bioprinting” refers to the process of creating three-dimensional (3D) structures by depositing living cells, and / or biomaterials, and / or growth factors layer by layer using additive manufacturing techniques. It involves the precise placement of biological materials (whether containing living cells or not), to generate complex tissues and organs that emulate the natural structure and functionality of living tissues / organs or plant base ingredients mixed or not mixed with cultivated cells to create complex fibers structure to mimic food and animal derived product.

[0016] The term “forming” (and its variants "formed", etc.) refers in an exemplary implementation to pumping, extruding, injecting, pouring, releasing, displacing, spotting, circulating, nebulizing, spraying, ink-jetting, jetting, or otherwise placing a fluid or material (e.g., fat or fat emulating material) in contact with another material (e.g., muscle, or muscle emulating material) using any suitable inkjet or other digital printing method. In an exemplary implementation, "forming" refers to the assembly of the 3D biostructure, optionally obtained with 3D files from 2D layer images derived from the 3D files, which, in another exemplary implementation are derived from various raster images (e.g., .dem) and / or vector data models.

[0017] In the context of the disclosure, the term “bio ink” or “bioink” means a liquid, semisolid, or solid composition optionally comprising a plurality of cells. In some exemplary implementations, bio-ink comprises cell solutions, cell aggregates, cell-comprising gels, multicellular bodies, or tissues. In some exemplary implementations, the bio-ink additionally comprises support material. In some exemplary implementations, the bio-ink additionally comprises non-cellular materials (e.g., cultivated food printing; cultivated cells mixed with plant base or plant based components only), which provide specific biomechanical properties that enable bioprinting.

[0018] Furthermore, in the context of the disclosure, the term "operable" means the system and / or the device and / or the program, or a certain element or step is fully functional, sized, adapted and calibrated, comprises elements for, has the functional interface, whether electric, mechanical, or chemical, and meets applicable operability requirements to perform a recited function when activated, coupled, implemented, actuated, effected, realized, contacted, or when an executable program isexecuted by at least one processor associated with the system and / or the device. In relation to systems and circuits, the term "operable" means the system and / or the circuit is fully functional and calibrated, comprises logic for, having the hardware and firmware necessary, as well as the circuitry for, and meets applicable operability requirements to perform a recited function when executed by at least one processor.

[0019] The term “slidably coupled” is used in its broadest sense to refer to elements that are coupled in a way that permits one element to slide or translate with respect to another element. Additionally, the term “slidably” or “slidably coupled” refers to movement of one surface (for example carrier frame 130) over a second surface (shelf sub-assembly 12 lip 1263) while maintaining smooth continuous contact between the two surfaces. In another embodiment, the term “slidably coupled” means a state in which two or more components are coupled to one another such that at least one of the components (e.g., carrier frame 130) at least slides with respect to another component (e.g., sliding spacers 132, 132’ (sec e.g., FIG.s IB, 1C)).

[0020] In the context of the disclosure, the term “drivetrain” refer to the group of components that deliver mechanical power from the prime power source to the driven components, parts, or assemblies, without reference to the power generator, such as a motor (e.g., servo motor, or an electric motor). Conversely, the term “powertrain” refers to those components that enable motion of one component relative to another, which does include to power source.

[0021] As used herein, the term “manifold” the term “manifold” or any variation thereof as used throughout the specification is defined hereinafter to include but is not limited to: a component for operating several devices of any kind at generally the same time; and also generally refers to a structure that is provided to assist in applying reduced pressure to, delivering fluids or materials to, or removing fluids to or from a given site. A manifold can typically include a plurality of flow channels or pathways. The plurality of flow channels may be interconnected to improve distribution of fluids or gas provided. Examples of manifolds may include, without limitation, devices that have structural elements arranged to form flow channels, cellular foam, such as open-cell foam, porous tissue collections, and liquids, gels, plant base materials, edible materials and foams that include or cure to include flow channels. The manifold material may also be a combination or layering of materials.

[0022] Here the term "outlet" includes any connector(s) for attaching to conduit associate with electrical, chemical, pneumatic, hydraulic, and the like subject matter.

[0023] In the context of the disclosure, the term “liquid communication” refers to a flow line (e.g. tubing or ducts) allowing for passage of liquid between two units, optionally through a number of intermediate units. A pressure source is in liquid communication with a filter for instance if the outlet for the filter is connected with an inlet of the mouth of a user and the like. Furthermore, as used herein, the term “liquid communication” means that liquid such as water is able to travel from one compartment or component to another compartment or component. The filter assembly can but need not include a liquid distribution layer, or the like, to channel or direct liquid to the other location(s)

[0024] Throughout bioprinting, it may be desired to maintain laminar flow of the bio-ink through certain components of the print head, while allowing for turbulent flow in others. In the context of the disclosure, “laminar flow” of bio-ink means stable, non-recirculating flow. In other words, there are no zones of recirculation, and turbulence is negligible. In each implementation the pipes, bores tubes and conduits defined in the various elements and components disclosed and claimed herein, arc each having dimensions, as is known to the art, allowing such non-turbulcnt flow under flow rates used. Certain configurations exemplified herein, are designed to be carried out in devices comprising pipes, bores tubes and conduits of a size such that the Reynolds number for flow within the channel is below about 2100, or is below about 10. Reynolds number is the ratio of inertia to apparent viscosity. Alternatively, “laminar flow” means pipes, bores tubes and conduits, which allow for laminar flow under hydrodynamic conditions, defined by viscosity and flow speed. For a pipes, bores tubes and conduits of given dimensions, laminar’ flow can be achieved for a liquid of certain viscosity and flow speed. The dimensions of the pipes, bores tubes and conduits are chosen such that laminar flow is maintained, i.e., low Reynold's number, preferably below about 1. That is, at least one dimension — either the diameter or the flow rate — is adapted sized and configured to maintain a low Reynold's number and hence laminar flow. In other exemplary implementations, for example, during cleaning procedures (such as clean-in-place (CIP), flow is typically turbulent).

[0025] As may also be used herein, the terms “central processing module” (CPM), “module”, “processing circuit”, and / or “processing unit” may be a single processing device or a plurality of processing devices. Such a processing device may be a microprocessor, micro-controller, digital signal processor, microcomputer, central processing unit, field programmable gate array, programmable logic device, state machine, logic circuitry, analog circuitry, digital circuitry, and / or any device that manipulates signals (analog and / or digital) based on hard coding of the circuitry and / or operational instructions (in other words, firmware). CPM (700, see e.g., FIG. 1 A), processing circuit,and / or processing unit may have an associated memory and / or an integrated memory element, which may be a single memory device, a plurality of memory devices, and / or embedded circuitry of the processing module, module, processing circuit, and / or processing unit. Such a memory device may be a read-only memory, random access memory, volatile memory, non-volatile memory, static memory, dynamic memory, flash memory, cache memory, and / or any device that stores digital information.

[0026] Furthermore, the term “processor” is defined as including, but not necessarily being limited to, an instruction execution system such as a computer / processor-based system, an Application Specific Integrated Circuit (ASIC), a computing device, or a hardware and / or software system that can fetch or obtain the logic from a non-transitory storage medium or a non-transitory computer- readable storage medium and execute the instructions contained therein. “Processor” can also include any controller, state-machine, microprocessor, cloud-based utility, service or feature, or any other analogue, digital and / or mechanical implementation thereof. In addition, the computer program (software and / or firmware), can comprise program code means for carrying out the steps to facilitate the bioprinting and control the print head and any of its components, for example, dispensing elements 14q described herein, as well as a computer program product comprising program code means stored on a medium that can be read by a computer, such as a hard disk, SATA CD-ROM, DVD, USB memory stick, or a storage medium that can be accessed via a data network, such as the Internet or Intranet, when the computer program product is loaded in the main memory of a computer and is carried out by the computer. Thus, the terms “non-transitory storage medium” and non-transitory computer-readable storage medium” are defined as including, but not necessarily being limited to, any media that can contain, store, or maintain programs, information, and data. Non-transitory storage medium and non-transitory computer-readable storage medium may include any one of many physical media such as, for example, electronic, magnetic, optical, electromagnetic, or semiconductor media.

[0027] Furthermore, various interconnection configurations ("topologies") for connecting multiple subsystems using one or more serial and / or parallel busses are used, as are the use of separate ("non-multiplexed") or shared ("multiplexed") address and data bus-lines. IN an exemplary implementation, the term "bus" will refer to a parallel, multiplexed or non-multiplexed bus in which subsystems connect through designated cable to a computerized control module.

[0028] The term "comprising" and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers,and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, "including", "having" and their derivatives.

[0029] All ranges disclosed herein are inclusive of the endpoints, and the endpoints are independently combinable with each other. “Combination” is inclusive of blends, mixtures, alloys, reaction products, and the like. The terms “a”, “an” and “the” herein do not denote a limitation of quantity, and are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The suffix “(s)” as used herein is intended to include both the singular and the plural of the term that it modifies, thereby including one or more of that term (e.g., the cell(s) includes one or more cells). Reference throughout the specification to “one exemplary implementation”, “another exemplary implementation”, “an exemplary implementation”, and its derivatives (implementation, configuration, examples, circumstances etc.) and so forth, when present, means that a particular element (e.g., feature, structure, and / or characteristic) described is included in at least one configuration described herein, and may or may not be present in other implementations. In addition, it is to be understood that the described elements may be combined in any suitable manner in the various examples, brought to provide clarifying examples.

[0030] All ranges disclosed herein are inclusive of the endpoints, and the endpoints are independently combinable with each other. Furthermore, the terms “first,” “second,” and the like, herein do not denote any order, quantity, or importance, but rather are used to denote one element from another...Further, as used herein, directional references such as "top", "base", "bottom", "end", "side", "interior", "exterior", "upper", "middle", "lower", "front" and "rear" do not limit the respective components to such orientation, but merely serve to distinguish these structural characteristics from one another.

[0031] Likewise, the term "about" means that amounts, sizes, formulations, parameters, and other quantities and characteristics are not and need not be exact, but may be approximate and / or larger or smaller, as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the ail. In general, an amount, size, formulation, parameter or other quantity or characteristic is "about" or "approximate" whether or not expressly stated to be such. For example, “about” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms ofdegree should be construed as including a deviation of at least ±5% or at least ±10% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0032] In an exemplary implementation, a 3D model of an edible product is formed by depositing bioink (or other support material) drop-by-drop. Each of bioink drop (interchangeable with bio ink), has to be dispensed on demand. To get a product with high accuracy a high printing resolution is needed, which calls for small, controllable drops, meaning, the drops need to be consistently of a specific size, which is no easy task. The print head assembly disclosed herein provides the ability to deposit a drop of the desired size at the right moment, in the precise location.

[0033] A more complete understanding of the assembly of print heads operable in a bioprinting systems for the fabrication of edible biostructures using drop-on-demand dispensers disclosed herein can be obtained by reference to the accompanying drawings. These figures (also referred to herein as “FIG.”) are merely schematic representations based on convenience and the ease of demonstrating examples of the present disclosure, and arc, therefore, not intended to indicate relative size and dimensions of the devices or components thereof, their relative size relationship and / or to define or limit the scope of the exemplary implementations illustrating them. Although specific terms are used in the following description for the sake of clarity, these terms are intended to refer only to the particular structure of the exemplary implementations selected for illustration in the drawings, and are not intended to define or limit the scope of the disclosure. In the drawings and the following description below, it is to be understood that like numeric designations refer to components of like function.

[0034] Turning now to FIG. 1A-1D illustrating printing station 10 for bioprinter comprising: printing assembly 10 comprising: shelf sub-assembly 12, back plate 11 defining longitudinal axis XLH', coupled to shelf sub-assembly 12, having powertrain operable to vertically translate (e.g., in the Z-direction) shelf sub-assembly 12; plurality of bio-ink buffer tanks 15m coupled to shelf sub assembly 12, each mthof plurality of bio-ink buffer tanks 15m operably coupled to group (e.g., I- IX, see e.g., FIG. IB, 1C) of print heads 14q, wherein each mthof plurality of bio-ink buffer tanks 15m is in communication with adjacent buffer tank 15m (see e.g., FIG. IB, 1C), and wherein first buffer tank is in further communication with first bio-ink reservoir 140; carrier frame 130 supporting array of plurality of print heads 14q divided into groups (e.g., I-IX, see e.g., FIG. IB, 1C), wherein carrier frame 130 defines longitudinal axis Xu30 (see e.g., FIG. 1A), and is coupled to shelf sub-assembly 12. Also illustrated is plurality of pressurized gas manifold pairs 147n, 147’n (see e.g., FIG. 1 A), coupledto corresponding group (e.g., I-IX, see e.g., FIG. IB, 1C), wherein the n!hmanifold of plurality of pressurized gas manifolds 147n, 147’n are each being in communication with corresponding half pair 145n, or 145’n of pressurized gas manifold of adjacent group, and wherein first pair of gas manifolds is in further liquid communication with first pressurized gas reservoir (not shown); conveyor 600 (see e.g., FIG.s 5A, 5B), operable to convey substrate 600f in parallel with longitudinal axis defined XLH by back plate 11; and central processing module (CPM) 700, in communication with back plate powertrain, plurality of bio-ink buffer tanks 15m, first compressed gas reservoir (not shown), and each of plurality of print heads 14q, CPM further comprising at least one processor in communication with non-transitory memory device storing thereon computer-readable medium with set of executable instructions configured, when executed by at least one processor to operate printing station.

[0035] As further illustrated in FIG.s 1A-1D, back plate 11 power train 112 comprises: motor 1120; and drivetrain coupled to motor 1120 and to rear side 120 of base plate 1300 (See e.g., FIG. 1A) of shelf sub-assembly 12, the drivetrain operable to vertically translate shelf sub-assembly 12 (in other words, in the Z-direction), while the drivetrain comprises: screw gear (or worm gear) 1121, operably coupled to motor 1120, screw gear (or worm gear) 1121 further comprises coupling member 1122 (see e.g., FIG. 1C) movably coupled to screw gear (or worm gear) 1121 and to rear side 120 of base plate 1300 of shelf sub-assembly 12; and pair of guide rails 113, 113’, coupled to back plate 11 front surface 111; and at least one carriage 114, 114’ slidably coupled to each guide rail 113, 113’, the at least one carriage 114, 114’, coupled to rear side 120 of base plate 1300 of shelf sub-assembly 12. In another exemplary implementation, each guide rail 113, 113’ has at least two carriages 114, 115 (114’, 115’) slidably coupled thereto.

[0036] Moreover, and as illustrated in FIG. IB, shelf sub-assembly 12 further comprises: pair of braces 126, 126’ disposed at opposite ends of front side 1301 of base plate 1300, each brace 126, 126’ having basal member 1261, 1261’ defining longitudinal axis XL1261 with at least one slot 1265d, 1265’d (see e.g., FIG. IB) in parallel with longitudinal axis XL1261,' and pair of sliding spacers 132, 132’ , each sliding spacer 132, 132’ operably coupled apically to basal member 1261, 1261’ of each brace 126, 126’, and to carrier frame 130 supporting array of plurality of print heads 14q. The pair of sliding spacers 132, 132’ configured to position longitudinal axis XL130 defined by carrier frame 130 supporting array of plurality of print heads 14q at predetermined angle a (sec e.g. FIG. 4) relative to longitudinal axis XL1261 defined in bottom surface of each of pair of braces, or relative to sample carrier substrate 600f.

[0037] As further illustrated in FIG.s 1 A-1C, print station 10 can further comprise several bus cards configured to operate the plurality of print heads 14q, divided into groups, and other sub-systems. As used herein, the term “bus card” is a reference to printed circuit boards with electronic components and a tab with a plurality of contacts that is received in the card slots of a data bus chassis. For example, first group bus, when employed, 123j is operable to control heating elements 1463, 14163’ and receive thermistor 1464, 1464’ data, transferred, in an exemplary implementation, using e.g., dedicated bus trunk to three groups bus card 122i, and from there to central control module 700. Likewise, group bus cards 124p, are operable to control switch valves 145, 145’ in each print head 14q.

[0038] In an exemplary implementation, control valves 159m controlling pressurized gas (e.g., pressurized air, at about 1 Atm. to 10 Atm., are likewise controlled by a separate dedicated processor 1590 (not shown), in communication with CPM 700.

[0039] In an exemplary implementation, and as illustrated in FIG.s 1 A-2B, each half pair of first gas manifold pairs 147n, 147’n comprises inlet port 1470 in liquid communication with first compressed gas reservoir (not shown), and plurality of outlet ports 147 In, 1471’n (see e.g., FIG.s 1C, ID) each outlet port 147 In, 1471’n in liquid communication with corresponding dispenser compressed gas inlet port 143, 143’ (see e.g., FIG. 2A) on each print head 14q. Likewise each mthof plurality of bio-ink buffer tanks 15m comprises: an inlet port 151 in liquid communication with the first bio-ink reservoir 140, and an outlet port 152, each inlet port 151 in liquid communication with a corresponding inlet port 151 on an adjacent bio-ink buffer tank 15m; and an outlet port 152, potentially in liquid communication with a first bio-ink recycling tank, or waste tank, (not shown), and wherein each outlet port 152 in liquid communication with a corresponding outlet port 152 on an adjacent bioink buffer tank 15m.

[0040] Turning for a moment to FIG. 3, illustrating X-Z cross section of bio-ink buffer tank 15m, comprising in an exemplary implementation, cylindrical body 155 defining cylindrical cavity 1550; cylindrical base 150, basally coupled to cylindrical body 155, cylindrical base defining plurality of apertures 155 lb, each bthaperture 1551b sized adapted and configured to be in communication with dispenser 146, 146’. Also shown is bio-ink inlet port 151, in liquid communication with cylindrical cavity 1550; bio-ink outlet port 152, in liquid communication with cylindrical cavity 1550; cylindrical cap 154, apically coupled to cylindrical body 155, cylindrical cap 154 being in liquid communication with second compressed gas reservoir 160 (not shown), via inlet port 1540. Moreover, cylindrical cap 154 further defining coaxial aperture 1541. Plunger comprised of stem 157 and cylindrical piston 156,with stem 157 extending through coaxial aperture 1541, and piston 156 sized and configured to be accommodated within cylindrical cavity 1550 defining in an exemplary implementation radial grooves 1560 configured to accommodate, for example, piston seals 1561, 1561’. Each mthof plurality of bio-ink buffer tanks 15m further comprises control valve 159m (see e.g., FIG. 1A) in communication with CPM 700, operable to actuate piston 156 at predetermined pressure (for example, between 4 atmospheres (Atm) and 6 Atm.), configured to bias bio-ink through each aperture 1551b defined in cylindrical base 150 to corresponding dispenser 146, 146’. As illustrated in FIG. 3, piston 156 defines chamber 1545, in liquid communication with compressed gas (e.g., compressed air) inlet port 1540, which is configured to bias piston 156 toward cylindrical base 150. Also illustrated, is convex basal surface 1562 of piston 156, which, in an exemplary implementation is complementary to concave surface 1505 of cylindrical base 150, such that full stroke of piston 156 is configured to completely empty cylindrical cavity 1550 from bio-ink.

[0041] Turning now to FIG.s 2A-2C, illustrating an exemplary implementation of the structure of print head 14q. As illustrated, each qthprint head 14q defines longitudinal axis Xu4q (see e.g., FIG. 2A) whereby each qthprint head 14q comprises pair of dispensers 146, 146’. As illustrated in FIG. 2A-2B, each qthprint head 14q further comprises; a housing 141 comprised of housing cover 1410 and housing base 1411, with upper surface 142 defining first pair of apertures 1430, 1430’, each aperture 1430, 1430’, (not marked) configured to accommodate corresponding inlet port 143, 143’ for compressed gas. Also shown, are second pair of exhaust apertures 144, 144’ configured to allow compressed gas to escape print head 14q upon actuation of dispensers 146, 146’. Pair of switch valves 145, 145’ are not shown, while their communication cable 1452, 1452’ is shown, each connection cable coupled to CPM 700, for example, through group bus 123j (see e.g., FIG. 1A).

[0042] As illustrated in FIG.s 2A, 2B, each qthprint head 14q further includes pair of dispensers 146, 146’, each dispenser 146, 146’ in liquid communication with corresponding switch valve 145, 145’, and corresponding compressed gas inlet port 143, 143’. Also illustrated are exhaust outlets 144, 144’. As further illustrated, and described further; FIG.s 2A-2B also illustrate mounting tabs 1412, 1412’ configured to couple each qth print head 14q to window 1330d defined in carrier frame 130 (see e.g. FIG. ID) defined by carrier frame 130 and transverse slats 133c. As indicated, communication cable 1452, 1452’ operable to communicate with CPM 700, through, for example group bus 123j, is also illustrated, as are leads 1463, 1463’ of heating element not shown, as well as thermistor entry apertures 1464, 1464’.

[0043] As further illustrated, each dispenser 146, 146’ further comprises upper unit isolator 1466, 1466’ and lower unit isolator 1465, 1465’ with heating element 1463, 1463’ (showing leads only exiting upper unit isolator 1466, 1466’ (see e.g., FIG. 2B)), while thermistor access aperture 1464, 1464’, is also shown. FIG. 2B also shows bio-ink inlet port 1453, 1453’ maintained in liquid communication with a corresponding bio-ink outlet 1500b of a corresponding group intermediate buffer tank 15m (see e.g., FIG.s IB, ID, 3).

[0044] Preventing the bio-ink 501 from getting out of dispenser 146, 146’ (through the nozzle 1460, 1460’) is achieved by gas pressure through compressed gas inlet 143, 143’ maintained at between about 3.5 Atm., and about 5.8 Atm. Actuating piston 156 using compressed gas through compressed gas inlet 1540 on each intermediate buffer tank 15m, is allowed to overcome the pressure exerted on each dispenser 146, 146’ through compressed gas inlet ports 143, 143’, allowing bio ink 501 to be expelled.

[0045] In addition, dispensers 146, 146’ is heated to keep environmental conditions as optimal as possible for the bio-ink. This means, among other things, keeping the temperature as close to the body temperature the selected cells will be exposed to in the biological sample. In addition, heating is beneficial since the viscosity of the bio-ink 501 is proportional to the temperature. Generally speaking, the higher the temperature, the lower is the viscosity of the bio-ink, resulting in smoother, laminar flow, given the narrow orifices the bio ink has to go through. In an exemplary implementation, as indicated, bio-ink 501 never comes in direct touch with heating element 1463, 1463’.

[0046] To print at least one layer, the set of executable instruction is configured, when executed to: using the array of the plurality of print heads 14q, actuate a first group of print heads to dispense the bio-ink 501 at a predetermined pattern, forming a first layer; elevate the shelf subassembly 12 to a predetermined height; and using the array of the plurality of print heads 14q, actuate a second group of print heads to dispense the bio-ink 501 at a predetermined pattern, forming a subsequent layer above the first layer.

[0047] As illustrated in FIG. 2C, the pair of nozzles 1460, 1460’ are configured to dispense the bio-ink 501 at a predetermined gap transverse to a printing direction. That predetermined gap can be between 1mm and 10 mm, selectable by changing the predetermined angle cr(see e.g., FIG. 4).

[0048] As further illustrated, the print head assembly has two main degrees of freedom, the print head assembly 10, moves across (Y direction) the conveyor 600, whereas the print tray 600f (see e.g., FIG.s 5A, 5B) provides the other degree of freedom by moving in the X direction with theconveyor 600. The movement of the print tray 600f can be infinite either by moving forward and backward, or by moving along a closed loop, and changing its longitudinal axis relative to the print head assembly 10. Along the X axis, whose length can be, for example, a few meters, several print head assemblies can be located. As the print tray 600f moves along this X axis located beneath the print head assembly 10. Whenever the print tray 600f is detected exactly beneath the proper sub-group of print heads 14q, printing is actuated.

[0049] To achieve high precision, on-demand deposition, the exact location in which deposition is to occur is calculated, then sent to a motion detection module (not shown) included with the print head station(s), in communication with CPM 700. The motion detection nodule 730 uses a high precision encoder 7300 (not shown), enabling the module (and operator) to know the location of the print tray 600f in real time with high accuracy. Once print tray 600f arrives at the deposition location, the motion detection module, sends the indication and deposition is performed by the print head 14q array group (c.g., I-IX, sec c.g., FIG.s IB, ID, 5A, 5B).

[0050] Formulating the bio-ink composition(s), may take into account the requirements, if any, imposed by the dispensing print head 14q and the surface characteristics (e.g., hydrophilic or hydrophobic, and the surface energy of and optionally provided substrate (print tray 600f), whether the desired flow is laminar or turbulent, differentiation constraints on stem cells, effect of organelles flow orientation / demixing and the like). For example, using dispensing elements 146, 146’ with at least one of: a piezo electric dispenser, a needle valve, a diaphragm, and a pinch valve, the dynamic viscosity of either the first bio-ink and / or the second bio-ink (measured at dispensing temperature temperature) can be, for example, between about 10 Poise (P) and about 10,000 P, or between about 50 P and about 7,500 P, for example, between about 100 P and about 5000 P at room temperature (23 °C - 25 °C).

[0051] For example, the first bio-ink 501 can comprise one or more of: a composition comprising: stem cells, edible plant based ingredients endothelial cells, muscle cells, fibroblast cells, mesothelial cells, pericyte cells, monocyte cells, plasma cells, mast cells, adipocyte cells, chondrocyte cells, cells population manipulated from a specific type of cell to another, or cells composition comprising one or more of the foregoing cells; a bio-compatible polymer; and optionally a photoinitiator.

[0052] Furthermore, the bio-ink in each print head 14q, can be the same or different. In other words, while bio-ink 501 in a first print head 14q can be, one or more of: a composition comprising:stem cells, edible plant based ingredients, endothelial cells, muscle cells, fibroblast cells, mesothelial cells, pericyte cells, monocyte cells, plasma cells, mast cells, adipocyte cells, chondrocyte cells, cells population manipulated from a specific type of cell to another, or cells composition comprising one or more of the foregoing cells; a bio-compatible polymer; and optionally a photoinitiator, bio-ink 501 circulated and dispensed through the at least one additional print head 14q, can be different, for example comprises: an extracellular matrix composition, PEGilated-methacrylate, chitosan, carrageenan, poly(lactic) acid, poly(lactic-co-glycolic acid), their methacrylate conjugates, copolymers, interpenetrating networks or a composition comprising one or more of the foregoing. In certain examples, the extracellular matrix (ECM) composition in the second bio-ink can comprise a composition configured to accelerate cell adhesion, promote cell colonization, proliferation, or manipulation, retain native growth of the cells, and / or organ and / or tissue. These can be, for example, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), bone morphogenic protein (BMP), insulin-likc growth factor (IGF), glucoscaminoglycan (GAG), Transforming growth factor (TGF) or extracellular matrix compound composition comprising the foregoing.

[0053] Methods utilizing print head 14q (see e.g., FIG. 4-5B), for use in the bioprinting or forming the 3D, cell-embedded biostructure (e.g., tissue, organ, tissue scaffold, connective tissue) can dispense (and derivatives thereof - understood to refer to any device or technique that deposits, dispenses, transfers or creates material on a surface in a controlled accretive or additive manner), can deposit the bio-ink droplet(s) upon demand, in other words, as a function of various process parameters such as a conveyor speed, desired cells layer thickness, layer type (e.g., certain type of cell or several types of cells, either manipulated (differentiated) or not ), location and the like.

[0054] The print head 14q can be used in an exemplary implementation to pattern the bio- ink(s) by expelling droplets of the liquid bio-ink 501 nozzle 1460, 1460’ one-at-a-time, as the print head 14q (or the substrate) is maneuvered, for example in two (X-Y, see e.g., FIG. 5A) (it should be understood that the dispensing print-head can also move in the Z axis) dimensions at a predetermined distance above the substrate (600f) or any subsequent layer. The height of the print head 14q nozzle 1460, 1460’ can be changed with the number of layers, maintaining for example a fixed distance. Each droplet can be configured to take a predetermined trajectory to substrate 600f on command by, for example a pressure impulse, or biasing element strength, via nozzle 1460, 1460’.

[0055] Moreover, a second bio-ink used in conjunction with a first bio-ink, can further comprise other additives that affect colonization, proliferation, adherence, inhibit apoptosis, or othermanipulation of the cells, retain native growth of the cells, and / or organ and / or tissue, crosslinking agents, and other similar functions as well as other plant-based ingredients. Accordingly and in an exemplary implementation, the second bio-ink used in the methods, systems and compositions for use in the direct inkjet bioprinting of a composite biostructure (e.g., tissue, organ, tissue scaffold, connective tissue), can further comprise: cells manipulation triggering compounds, (for example, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), bone morphogenic protein (BMP), insulin-like growth factor (IGF), glucoseaminoglycan (GAG), Transforming growth factor (TGF) or signaling compound composition comprising the foregoing), as well as therapeutically effective compounds, antimicrobial compounds, immunosuppressing compounds and the like.

[0056] Accordingly, and in an exemplary implementation, provided herein is a printing station for a bioprinter, or a liquid delivery system comprising: a shelf sub-assembly, a back plate defining a longitudinal axis; coupled to the shelf sub-assembly, having a powertrain operable to vertically translate the shelf sub-assembly; a plurality of bio-ink buffer tanks coupled to the shelf sub assembly, each of the plurality of bio-ink buffer tanks operably coupled to a group of print heads, wherein each of the plurality of bio-ink buffer tanks is in communication with an adjacent buffer tank, and wherein a first buffer tank is in further communication with a first bio-ink reservoir; a frame supporting an array of a plurality of print heads divided into the groups, wherein the frame defines a longitudinal axis and is coupled to the shelf sub-assembly; a plurality of pressurized gas manifold pairs, coupled to a corresponding group, wherein each of the plurality of pressurized gas manifolds are each being in communication with a corresponding half pair of the pressurized gas manifold of an adjacent group, and wherein a first pair of gas manifolds is in further liquid communication with a first pressurized gas reservoir; a conveyor, operable to convey a substrate in parallel with the longitudinal axis defined by the back plate; and a central processing module (CPM), in communication with the back plate powertrain, the plurality of bio-ink buffer tanks, the first compressed gas reservoir, and each of the plurality of print heads, the CPM further comprising at least one processor in communication with a non-transitory memory device storing thereon a computer-readable medium with a set of executable instructions configured, when executed by the at least one processor to operate the printing station, wherein (i) the back plate power train comprises: a motor (e.g., servo motor, or an electric motor); and a drivetrain coupled to the motor as well as to a rear side of a base plate of the shelf sub-station, the drivetrain operable to vertically translate (in other words along a Z-axis) the shelf sub-station, (ii) the drivetrain comprises: a screwgear, or worm gear, operably coupled to the motor, the screw (or worm) gear further comprises a coupling member movably coupled to the screw gear and to the rear side of the base plate of the shelf sub-station; and a pair of guide rails, coupled to the back plate; and at least one carriage slidably coupled to each guide rail, the at least one carriage coupled to the rear side of the base plate of the shelf sub-station, wherein (iii) the shelf sub-station further comprises: a pair of braces disposed at opposite ends of the front side of the base plate, each having a bottom surface defining a longitudinal axis with at least one slot in parallel with the longitudinal axis; and a pair of braces (or support members), each brace coupled to a sliding spacer; the pair of sliding spacer being operably coupled to the bottom surface of each brace, as well as to the frame supporting the array of the plurality of print heads, the pair of sliding spacers configured to position the longitudinal axis defined by the frame supporting the array of a plurality of print heads at a predetermined angle relative to the longitudinal axis defined in the bottom surface of each of the pair of braces, wherein(iv) each print head defines a longitudinal axis, and further comprises a pair of dispensers, whereby(v) each bio-ink buffer tank further comprises: a cylindrical body defining a cylindrical cavity; a cylindrical base, basally coupled to the cylindrical body, the cylindrical base defining a plurality of apertures, each aperture sized adapted and configured to be in communication with a dispenser; a bio-ink inlet port, in liquid communication with the cylindrical cavity; a bio-ink outlet port, in liquid communication with the cylindrical cavity; a cylindrical cap, apically coupled to the cylindrical body, the cylindrical cap being in liquid communication with a second compressed gas reservoir, the cylindrical cap further defining a coaxial aperture; a plunger comprising a stem and a cylindrical piston, sized and configured to be accommodated within the cylindrical cavity; and a control valve in communication with the CPM, operable to actuate the piston at a predetermined pressure, configured to bias the bio-ink through each aperture defined in the cylindrical base to the corresponding dispenser, wherein (vi) each half pair of the first gas manifold pairs comprises an inlet port in liquid communication with the first compressed gas reservoir, and a plurality of outlet ports, each outlet port in liquid communication with a corresponding dispenser compressed gas inlet port on each print head, wherein (vii) each bio-ink buffer tank comprises: an inlet port in liquid communication with the first bio-ink reservoir, and an outlet port, each inlet port in liquid communication with a corresponding inlet port on an adjacent bio-ink buffer tank; and an outlet port, potentially in liquid communication with a first bio-ink recycling tank, or waste tank, and wherein each outlet port in liquid communication with a corresponding outlet port on an adjacentbio-ink buffer tank, wherein (viii) the carrier frame supporting the array of a plurality of print heads defines a plurality of windows, adapted sized and configured to accommodate the group of the plurality of print heads, (ix) each print head further comprises: a housing, comprised of a housing cover and a housing base, wherein the hosing cover having an upper surface defining a first pair of pressurized gas apertures, each aperture configured to couple to a corresponding outlet in the plurality of pressurized gas manifold pairs; a pair of switch valves (referring to a device that controls the flow of fluid by electronically or mechanically switching between different fluid sources, or switching between dispensing and non-dispensing modes, allowing for the selection of different fluids or the start / stop of fluid flow, thereby enabling precise control over the dispensing operations), each switch valve being in liquid communication with a corresponding pressurized gas inlet port; a pair of dispensers, each dispenser being in further liquid communication with a corresponding switch valve, and a corresponding bio-ink inlet port, (x) each dispenser further comprises: an upper unit isolator; a lower unit isolator; a flanged nozzle, coupled to a nozzle bolt; and the nozzle bolt itself, (xi) each dispenser further comprises a heating element in communication with the CPM, (xii) each heating element is operably coupled to a thermistor in communication with the CPM, wherein (xiii) wherein the pair of flanged nozzle are configured to dispense the bio-ink at a predetermined gap transverse to a printing direction, (xiv) the electric heating body is in communication with the CPM, wherein (xv) the gap between the bio-ink drops dispensed by each nozzle is selectable by changing the predetermined angle (of the substrate on the conveyor e.g.), (xvi) the predetermined gap being between 1mm and about 10 mm, wherein (xvii) the set of executable instruction is configured, when executed to: using the plurality of print heads prearranged in an array, actuate a first group of print heads to dispense the bio-ink at a predetermined pattern, forming a first layer; then elevate the shelf sub-station in the Z-axis direction, to a predetermined height above that layer; and using the array of the plurality of print heads, actuate a second group of print heads to dispense the bio-ink at a (different or the same) predetermined pattern, forming a subsequent layer above the first layer, wherein (xviii) the printing apparatus (interchangeable with “station”, “system”, “assembly” or “subassembly”), or additionally or alternatively, (xix) nine groups of 4 print heads (in other words 72 dispensing nozzles), wherein each heating element in each group (of the nine), and each thermistor is in communication with a second group bus disposed on a second panel portion coupled to the shelf sub-assembly, the second group bus being in communication with the CPM.

[0057] In another exemplary implementation, provided herein is a bioprinter comprising: a plurality of the printing station of any one of claims 1-19; and a central command and control module (C3M) operable to control the plurality of printing stations

[0058] Although the foregoing disclosure for print head assembly been described in terms of some exemplary configurations, other configurations will be apparent to those of ordinary skill in the art from the disclosure herein. Indeed, the novel systems, devices, and assemblies described herein may be embodied in a variety of other forms without departing from the spirit thereof. Accordingly, other combinations, omissions, substitutions and modifications will be apparent to the skilled artisan in view of the disclosure herein.

Claims

What is claimed:

1. A printing station for a bioprinter comprising: a) a shelf sub-assembly, b) a back plate defining a longitudinal axis; coupled to the shelf sub-assembly, having a powertrain operable to vertically translate the shelf sub-assembly; c) a plurality of bio-ink buffer tanks coupled to the shelf sub assembly, each of the plurality of bio-ink buffer tanks operably coupled to a group of print heads, wherein each of the plurality of bio-ink buffer tanks is in communication with an adjacent buffer tank, and wherein a first buffer tank is in further communication with a first bio-ink reservoir; d) a frame supporting an array of a plurality of print heads divided into the groups, wherein the frame defines a longitudinal axis and is coupled to the shelf sub-assembly; e) a plurality of pressurized gas manifold pairs, coupled to a corresponding group, wherein each of the plurality of pressurized gas manifolds are each being in communication with a corresponding half pair of the pressurized gas manifold of an adjacent group, and wherein a first pair of gas manifolds is in further liquid communication with a first pressurized gas reservoir; f) a conveyor, operable to convey a substrate in parallel with the longitudinal axis defined by the back plate; and g) a central processing module (CPM), in communication with the back plate powertrain, the plurality of bio-ink buffer tanks, the first compressed gas reservoir, and each of the plurality of print heads, the CPM further comprising at least one processor in communication with a non-transitory memory device storing thereon a computer-readable medium with a set of executable instructions configured, when executed by the at least one processor to operate the printing station.2 The printing station of claim 1, wherein the back plate power train comprises: a) a motor; and b) a drivetrain coupled to the motor and to a rear side of a base plate of the shelf substation, the drivetrain operable to vertically translate the shelf sub-station.3 The printing station of claim 2, wherein the drivetrain comprises: a) a screw gear, operably coupled to the motor, the screw gear further comprises a coupling member movably coupled to the screw gear and to the rear side of the base plate of the shelf sub-station; and b) a pair of guide rails, coupled to the back plate; andc) at least one carriage slidably coupled to each guide rail, the at least one carriage coupled to the rear side of the base plate of the shelf sub- station.

4. The printing station of claim 1, wherein the shelf sub-station further comprises: a) a pair of braces disposed at opposite ends of the front side of the base plate, each having a bottom surface defining a longitudinal axis with at least one slot in parallel with the longitudinal axis; and b) a pair of braces, each brace coupled to a sliding spacer; and c) the pair of sliding spacer operably coupled to the bottom surface of each brace, and to the frame supporting the array of the plurality of print heads, the pair of sliding spacers configured to position the longitudinal axis defined by the frame supporting the array of a plurality of print heads at a predetermined angle relative to the longitudinal axis defined in the bottom surface of each of the pair of braces.5 The printing station of claim 4, wherein each print head defines a longitudinal axis, each print head comprises a pair of dispensers.6 The printing station of claim 5, wherein each bio-ink buffer tank further comprises: a) A cylindrical body defining a cylindrical cavity; b) A cylindrical base, basally coupled to the cylindrical body, the cylindrical base defining a plurality of apertures, each aperture sized adapted and configured to be in communication with a dispenser; c) A bio-ink inlet port, in liquid communication with the cylindrical cavity; d) A bio-ink outlet port, in liquid communication with the cylindrical cavity; e) A cylindrical cap, apically coupled to the cylindrical body, the cylindrical cap being in liquid communication with a second compressed gas reservoir, the cylindrical cap further defining a coaxial aperture; f) A plunger comprising a stem and a cylindrical piston, sized and configured to be accommodated within the cylindrical cavity; and g) A control valve in communication with the CPM, operable to actuate the piston at a predetermined pressure, configured to bias the bio-ink through each aperture defined in the cylindrical base to the corresponding dispenser.7 The printing station of claim 5, wherein each half pair of the first gas manifold pairs comprises an inlet port in liquid communication with the first compressed gas reservoir, and aplurality of outlet ports, each outlet port in liquid communication with a corresponding dispenser compressed gas inlet port on each print head.

8. The printing station of claim 6, wherein each bio-ink buffer tank comprises: a) an inlet port in liquid communication with the first bio-ink reservoir, and an outlet port, each inlet port in liquid communication with a corresponding inlet port on an adjacent bio-ink buffer tank; and b) an outlet port, potentially in liquid communication with a first bio-ink recycling tank, or waste tank, and wherein each outlet port in liquid communication with a corresponding outlet port on an adjacent bio-ink buffer tank.9 The printing station of claim 5, wherein the carrier frame supporting the array of a plurality of print heads defines a plurality of windows, adapted sized and configured to accommodate the group of the plurality of print heads.10 The printing station of claim 7, wherein each print head further comprises: a) a housing, comprised of a housing cover and a housing base, wherein the hosing cover having an upper surface defining a first pair of pressurized gas apertures, each aperture configured to couple to a corresponding outlet in the plurality of pressurized gas manifold pairs; b) a pair of switch valves, each switch valve in liquid communication with a corresponding pressurized gas inlet port; c) a pair of dispensers, each dispenser in liquid communication with a corresponding switch valve, and a corresponding bio-ink inlet port.11 The printing station of claim 9, wherein each dispenser comprises: a) an upper unit isolator; b) a lower unit isolator; c) a flanged nozzle, coupled to a nozzle bolt; and d) the nozzle bolt.12 The printing station of claim 10, wherein each dispenser further comprises a heating element in communication with the CPM.13 The printing station of claim 11, wherein each heating element is operably coupled to a thermistor in communication with the CPM,14 The printing station of claim 11, wherein the pair of flanged nozzle arc configured to dispense the bio-ink at a predetermined gap transverse to a printing direction.

15. The printing station of claim 11, wherein the electric heating body is in communication with the CPM.

16. The printing station of claim 14, wherein the gap between the bio-ink drops dispensed by each nozzle is selectable by changing the predetermined angle.

17. The printing station of claim 15, wherein the predetermined gap is between 1mm and 10 mm.

18. The printing station of claim 16, wherein the set of executable instruction is configured, when executed to: a) using the array of the plurality of print heads, actuate a first group of print heads to dispense the bio-ink at a predetermined pattern, forming a first layer; b) elevate the shelf sub-station to a predetermined height; and c) using the array of the plurality of print heads, actuate a second group of print heads to dispense the bio-ink at a predetermined pattern, forming a subsequent layer above the first layer.

19. The printing station of claim 12, further comprising nine groups of 4 print heads, wherein each switch valve in each group, and each bio-ink buffer tank control valve is in communication with a first group bus disposed on a first panel portion coupled to the shelf sub-assembly, the first group bus being in communication with the CPM.

20. The printing station of claim 12, further comprising nine groups of 4 print heads, wherein each heating element in each group, and each thermistor is in communication with a second group bus disposed on a second panel portion coupled to the shelf sub-assembly, the second group bus being in communication with the CPM.

21. A bio printer comprising: a plurality of the printing station of any one of claims 1-19; and a central command and control module (C3M) operable to control the plurality of printing stations.

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