Dielectric spectroscopy device containing systems, dielectric spectroscopy devices, and methods thereof

Abstract

The present disclosure is directed to a system for collecting or extruding a cell-containing biological material. The system includes an extrusion / collection device comprising: an orifice and a chamber operably connected to the orifice and a dielectric spectroscopy (DS) device operably connected to the extrusion / collection device. The DS device monitors biologically relevant attributes of a cell-containing biological material as it is extruded from or collected into the extrusion / collection device. Also disclosed are methods of using the system disclosed herein.

Description

DIELECTRIC SPECTROSCOPY DEVICE CONTAINING SYSTEMS, DIELECTRIC SPECTROSCOPY DEVICES, AND METHODS THEREOF

[0001] This application claims the priority benefit of U.S. Provisional Patent Application Serial No. 63 / 614,480, filed December 22, 2023, which is hereby incorporated by reference in its entirety.FIELD

[0002] The present disclosure relates to dielectric spectroscopy (DS) device-containing systems capable of monitoring biologically relevant attributes of cell-containing biological material, DS devices, and methods thereof.BACKGROUND

[0003] A variety of high-end cell processing techniques require the mass transportation of cells from one area to another. Examples include cell therapies, bioprinting, cell manufacturing, patient sample collection, etc. In each of these cases, cell-laden samples are assessed for quality control post-transfer. Separate sampling and assessment creates a significant time delay in the manufacturing or treatment process and increases the unit operations necessary for the manufacturing process. As such, there is a need for real-time measurement of cell health, such as cell concentration, during transfer. Currently, limited techniques exist for this application. Optical spectroscopy can non-destructively detect cell concentration but struggles with sensitivity to high cell concentrations and measuring moving samples with flow rate. Flow cytometry functions under flow rate and gives accurate information on a cell-by-cell basis but is too slow for real-time assessment of samples with high concentrations (e.g., 104- 105cells / mL), failing to assess the bulk properties of a sample.

[0004] It is important to measure a wide range of cell concentrations for transfer processes in which the concentration of the sample has the potential to vary greatly from the target value. Currently, there are no commercially available products that detect a wide range of cell concentrations during flow conditions. All available literature and products assess cellular properties after bioprinting the constructs using techniques such as dielectric spectroscopy, oxygen consumption, and ultrasound. Further, there are no commercially available products that detect cell concentration in real-time during transfer processes. Available literature and products use sensors and non-destructive techniques while the sample is stationary in a container such as aflask or bioreactor. Examples of current techniques include, but are not limited to, oxygen consumption, optical spectroscopy, ultrasound, and dielectric spectroscopy.

[0005] As technology for scaling up cell manufacturing improves, the need for real-time monitoring of cell concentration increases. Cell samples are often times transferred from a source location, such as patients, bioreactors, or flasks, to a destination such as a container, bioreactor, or bioprinter. In addition, a single cell sample may be transferred several times during cell manufacturing; for instance, a sample collected from a patient for cell therapy will be cultured, expanded, re-packaged, transported, and injected back into the patient. To assure that quality control standards are met, portions of the sample are externally assessed posttransfer, which creates a significant time delay in the manufacturing process. Therefore, there is a need for real-time measurement of cell concentration during sample transfer.

[0006] Due to the wide range of cell concentrations collected from source locations, evaluation of cell concentration on the order of 104- 109cells / mL during flow conditions from 10’ O3mL / min is necessary for real-time transfer analysis. Currently, several technologies have been investigated for detection of cell health in suspension, well-plates, bioreactors, or flasks, but there is no method for measuring bulk cellular properties with flow conditions. Flow cytometry is commonly used to assess samples on a cell-by-cell basis, but it is too slow for real-time assessment of samples with high concentration >104cells / mL, failing to assess the bulk properties of a sample.

[0007] To address the issue of real-time assessment of cell-laden samples, a device was previously created (see U.S. Patent No. 12,019,040) to measure cell concentration in real-time for bioprinting. This device used a single electrode pair to detect cellular properties on the order of 106cells / mL and was sensitive to cell concentrations from 25 x 106- 125 x 106cells / mL. However, the device was not sensitive to lower cell concentrations.

[0008] Another device used copper wire embedded into a syringe tip to assess cellular properties. However, this device was limited to cell concentrations at 5 x 105cells / mL. In addition, cell-laden samples were not tested under flow rate (Haring et al., “3D Bioprinting Using Hollow Multifunctional Fiber Impedimetric Sensors,” Biofabrication 12(3):035026 (2020)). A further device utilized an impedance-based sensor for detection of cell growth in CAR-T cell manufacturing. This device uses electrodes sandwiched between PDMS (Liu et al., “A Disposable Impedance-based Sensor for In-Line Cell Growth Monitoring in CAR-T Cell Manufacturing” Bioelectrochemistry 152: 108416 (2023)).

[0009] Additionally, one study assessed bioprinted constructs post-fabrication using dielectric spectroscopy. The investigators used a dielectric spectroscopy probe in a dishcontaining fluid and a bioprinted construct to report dielectric parameters of various cell types. The samples were not assessed during flow conditions and only one cell concentration (2xl06cells / construct) was investigated (Shohan et al., “Non-Destructive Quality Monitoring of 3D Printed Tissue Scaffolds via Dielectric Impedance Spectroscopy and Supervised Machine Learning,” Procedia Manufacturing 53:636-643 (2021)).

[0010] The present disclosure is directed to overcoming these and other deficiencies in the art.SUMMARY

[0011] One aspect of the present disclosure relates to a system for collecting or extruding a cell-containing biological material. The system includes an extrusion / collection device comprising an orifice and a chamber operably connected to the orifice. A dielectric spectroscopy (DS) device is operably connected to the extrusion / collection device. The DS device comprises at least two sets of electrodes, each set comprising a plurality of electrodes. A first set of the at least two sets of electrodes comprises a first material and a first geometrical configuration and a second set of the at least two sets of electrodes comprise a second material and a second geometrical configuration. At least one of the first and second material and the first and second geometrical configuration differ from each other. The DS device is capable of monitoring biologically relevant attributes of the cell -containing biological material as the cell -containing biological material is extruded from or collected into the extrusion / collection device.

[0012] Another aspect of the present disclosure relates to a method of depositing a cellcontaining biological material. This method involves providing a system disclosed herein; extruding from an extrusion device of the system a cell -containing biological material; and monitoring biologically relevant attributes of the cell-containing biological material as the cellcontaining biological material is extruded from the device.

[0013] A further aspect of the present disclosure relates to a method of collecting a cellcontaining biological material. This method involves providing a system disclosed herein; collecting into a collection device of the system a cell-containing biological material; and monitoring biologically relevant attributes of the cell-containing biological material as the cellcontaining biological material is collected into the device.

[0014] Another aspect of the present disclosure relates to a system for collecting or extruding a cell-containing biological material. The system includes a syringe device comprising a housing having opposed distal and proximal ends, where the distal end comprises an orifice. The syringe device also includes a fitting connected at the orifice for interchangeably connectinga needle, a plunger axially movable within the housing between an advanced position near the distal end and a retracted position near the proximal end, and an elongate rod being connected to the plunger to move the plunger axially between the advanced and retracted positions and extending through the proximal end of the housing. The system also includes a dielectric spectroscopy (DS) device operably connected to the syringe device at the fitting. The DS device comprises a first set of electrodes and a second set of electrodes, the first set of electrodes comprises a first material and a first geometrical configuration, and the second set electrodes comprises a second material and a second geometrical configuration. At least one of the first and second materials and the first and second geometrical configurations differ from each other. The electrodes create an electrical field for detecting impedance of a cell -containing biological material as the cell -containing biological material is extruded from or collected into the syringe device.

[0015] A further aspect of the present disclosure relates to a dielectric spectroscopy (DS) device connectable to an extrusion / collection device, where the DS device comprises a first and second set of anti-parallel electrodes. The first set of anti-parallel electrodes comprises a first material and a first geometrical configuration and the second set of anti-parallel electrodes comprises a second material and a second geometrical configuration. At least one of the first and second material and the first and second geometrical configuration differ from each other. The first and second set of anti-parallel electrodes are configured to create an electrical field around a fluid passageway for detecting impedance of a cell -containing biological material as the cellcontaining biological material moves through the fluid passageway.

[0016] Another aspect of the present disclosure relates to an extrusion / collection device comprising an orifice and a chamber operably connected to the orifice. The extrusion / collection device is configured to be operably connected to a dielectric spectroscopy (DS) device comprising a first and a second set of electrodes, the first set of electrodes comprising a first material and a first geometrical configuration, the second set of electrodes comprising a second material and a second geometrical configuration. At least one of the first and second material and the first and second geometrical configuration differ from each other. The DS device is configured to monitor biologically relevant attributes of a cell-containing biological material as the cell-containing biological material is extruded from or collected into the extrusion / collection device.

[0017] A further aspect of the present disclosure relates to a system for measuring a parameter of a biological material. The system includes a material receiving device configured to contact the biological material, the material receiving device comprising an orifice and achamber operably connected to the orifice, the chamber being configured to receive the biological material therein. The system also includes a dielectric spectroscopy (DS) device comprising an electrode operably connected to the material receiving device. The DS device is configured to monitor a parameter of the biological material when the biological material is received in the material receiving device.

[0018] The systems and devices of the present disclosure detect a vastly greater range of cell concentrations (e.g., and without limitation, 104to 109cells / mL) with a higher degree of sensitivity by implementing a multi-electrode system. Accordingly, the multiple electrodes of the devices and systems vary in orientation, shape, size, and / or material to determine dielectric properties of a suspension with or without cells. Cell concentration is determined using data collected at particular frequencies with or without flow conditions. Further, for the systems and devices described in this disclosure, it was discovered that combining multiple electrodes with different geometries and materials can combat challenges in sensitivity, vastly improving upon the measurable range of cell concentrations. Additionally, the devices and systems of the present disclosure are able to reproducibly detect cell concentrations on the order of 104to 109cells / mL with and without flow rate.

[0019] Electrodes and devices described herein are integrated into various form factors for in-line detection of cell health, such as cell concentration, cell viability, and sterility for realtime monitoring during manufacturing processes (cell manufacturing, cell transfer, patient sample collection, etc.).

[0020] The present disclosure uses embedded or inserted electrodes to generate an electric field through the path of sample flow for in-line detection of cellular properties. Notably, the devices and systems of the present disclosure can detect cell concentrations within a range of e.g., 104to 109cells / mL or any value / range therein, in real-time under flow rates from 10'1- 103mL / min, or any value / range therein, allowing for rapid measurement of samples during transfer. Accordingly, the devices and systems of the present disclosure are versatile and adaptable to many fields such as cell therapy, cell manufacturing, and bioprinting with a focus on in-line measurement of cell concentration during sample transfer.

[0021] Existing bioprinting technology uses destructive techniques for analysis of constructs post-fabrication. The systems and devices described herein can reduce the reliance on destructive techniques by providing information on construct cellular properties in real-time.BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIGS. 1 A-1B depict one embodiment of a multi-electrode prototype, combining gold rod-shaped electrodes with platinum curved electrodes into a single chamber. FIG. 1 A shows a top-view with the gold rod electrodes on the surface. FIG. IB shows a bottom view with the gold rod electrodes obscured by the platinum curved electrodes.

[0023] FIG. 2 shows gold rod electrodes before and after heat shrink was added.

[0024] FIG. 3 depicts a platinum-coated curved electrode positioned in a chamber / housing unit with clamps holding the platinum electrode unit upright.

[0025] FIGS. 4A-4B are various illustrated views of embodiments of an electrode prototype. FIG. 4A shows a zoomed top view of one embodiment of a gold rod-shaped electrode prototype. FIG. 4B depicts that the distance between electrodes can vary.

[0026] FIGS. 5A-5B depict example illustrations of embodiments of a prototype device using platinum curved electrodes. FIG. 5A is a perspective view showing a platinum electrode device that is unattached. FIG. 5B is a top view showing a platinum electrode device that is attached to a commercial syringe and syringe tip.

[0027] FIG. 6 depicts various illustrated top views of embodiments of a gold rod electrode device connected to commercial syringes and syringe tips.

[0028] FIG. 7 shows a cross-section of one embodiment showcasing three electrode pairs with varied geometry using different conductive materials for each of the electrode pairs.

[0029] FIG. 8 is a graph showing that impedance measurements from platinum curved electrodes increased at low cell concentrations but plateaued at higher concentrations. Meanwhile, gold, rod-shaped electrodes showed impedance plateaued at low cell concentrations but increased at higher concentrations.

[0030] FIG. 9 is a graph showing the sensitivity of each electrode type displayed as the slope of cell concentration over the impedance of the sample. Sensitivity for gold rod electrodes according to one embodiment was highest at concentrations >1 x 106, and platinum curved electrodes according to one embodiment were most sensitive at <10 x 106cells / mL. Plateau regions for each electrode represent minimal sensitivity to changes in cell concentration using impedance.

[0031] FIG. 10 is a graph showing that the prototype displayed repeatable results after 5 independent experiments with n=3 per experiment. Data shown at 25 kHz. Notably, this trend implies that one embodiment of the device is capable of detecting cell concentrations greater than 125 x 106cells / mL.

[0032] FIG. 11 is a graph showing that the phase angle increased slightly with cell concentration at 25 kHz, indicating an increase in the inductive behavior of the sample. This trend is also noted in FIG. 9.

[0033] FIG. 12 is a graph showing device sensitivity for one embodiment of a gold, rodshaped electrode prototype and indicates that the greatest change in impedance with respect to cell concentration was from 10-100 kHz.

[0034] FIG. 13 is a graph showing device sensitivity for the gold, rod-shaped electrode prototype and indicates that the greatest change in phase angle with respect to cell concentration was from 1-10 kHz.

[0035] FIG. 14 is an illustration of a cross-section of one embodiment of an example device, showing gold rod electrodes in the path of sample flow during a transfer process from a syringe to another container.

[0036] FIG. 15 is a graph showing sensitivity of impedance (Z) to changes in cell concentration under flow at 2 mL / min. Sample concentration was altered approximately every 2 minutes. Sample impedance shifted in real-time in response to changes in bulk sample cell concentration. Regions of sample transition were characterized by an initial drop in impedance followed by a steady increase until reaching a steady value, indicating that the sample showed consistent concentration.

[0037] FIGS. 16A-16B are graphs depicting the stability of impedance (Z) and phase angle (9) as measurements under common flow rates for cell transfer. FIG. 16A shows that after normalizing to t=0 seconds, no significant difference was found in impedance or phase angle over 240 seconds, or 4 minutes, of continuous flow at 0.5, 2, or 4 mL / min with concentration at 25 x 106cells / mL. FIG. 16B shows that no significant difference was noted for other examined cell concentrations for 90 seconds of continuous flow at 2 mL / min.

[0038] FIGS. 17A-17B are illustrations of example form factors. FIG. 17A depicts transfer from syringe to bioprinter bed. FIG. 17B shows transfer from syringe pump to tube or container.

[0039] FIG. 18 is a depiction of various form factors that the present technology could be incorporated into for in-line assessment of cell concentration during sample transfer.

[0040] FIG. 19 is an illustration of one embodiment of a multi-electrode device containing two or more electrode types. The model shown features a pair of gold rod electrodes and curved platinum electrodes in series.

[0041] FIGS. 20A-20B are graphs showing the analysis of multi-electrode measurements using normalized impedance. FIG. 20A shows the data for the gold rod electrodes. FIG. 20Bshows the data for the curved platinum electrodes. The boxes indicate regions of high sensitivity to cell concentration. 60 kHz- 1,000 kHz constitutes electrode crosstalk.

[0042] FIGS. 21 A-21D are graphs showing impedance at various cell concentrations using specific frequencies. Connecting lines indicate sample average. FIG. 21 A shows that 0.1 kHz shows some resolution for the curved platinum electrodes with increasing impedance from 0.6-1.1 x 106cells / mL as well as from 79-112 x 106cells / mL. Gold rod electrodes were mostly unresponsive. FIG. 21B shows that similar to 0.1 kHz, 1 kHz showed changing impedance as cell concentration increased for the curved platinum electrodes but little change in behavior for gold rod electrodes. FIG. 21C shows that at 10 kHz, gold rod electrodes increased with increasing cell concentration, particularly from 28-112 x 106cells / mL. Meanwhile the curved platinum electrodes showed less impedance than shown at 0.1 and 1 kHz. FIG. 2 ID shows that electrode cross-talk results in sporadic and unstable measurements from both electrodes.

[0043] FIGS. 22A-22D are graphs showing multi -el ectrode dataset fitted to a piecewise, weighted linear regression. The y-axis displays Zceii. FIG. 22A shows that at 0.1 kHz, curved platinum electrodes show increasing impedance with increasing cell concentration. Gold rod electrodes show the opposite behavior. FIG. 22B shows that similar behavior is shown at 1 kHz compared to 0.1 kHz with opposite behaviors seen between the curved platinum and gold rod electrodes. FIG. 22C shows that at 10 kHz, the gold rod electrodes show increasing impedance with increasing cell concentration from 28-112 x 106cells / mL. Curved platinum electrodes also show increasing impedance, particularly from 0.6- 1.1 x 106cells / mL. FIG. 22D shows that crosstalk between both electrodes greatly impacted the collected data.DETAILED DESCRIPTIONDefinitions

[0044] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present disclosure herein described for which they are suitable as would be understood by a person skilled in the art. Before the present system, devices, and methods are described, it is to be understood that this disclosure is not limited to the particular systems, devices, or methodologies described, as these may vary. It is also to be understood that the terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of embodiments herein which will be limited only by the appended claims. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments described or shown herein, thepreferred methods, devices, systems, and materials are now described. All publications mentioned herein are incorporated by reference in their entirety. Nothing herein is to be construed as an admission that embodiments herein are not entitled to antedate such disclosure by virtue of prior invention.

[0045] Singular forms “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to “a method” includes one or more methods, and / or steps of the type described herein and / or which will become apparent to those persons skilled in the art upon reading this disclosure.

[0046] The term “about” or “approximately” includes being within a statistically meaningful range of a value. Such a range can be within an order of magnitude, such as within 50%, or within 20%, or within 10%, or within 5% (or any amount or range within 5-50%) of a given value or range. The allowable variation encompassed by the term “about” or “approximately” may depend on the context.

[0047] The term “and / or” as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that “at least one of’ or “one or more” of the listed items is used or present.

[0048] As will be understood by a person of ordinary skill in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof, as well as any value within a range. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, and so on. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, and so on. As will also be understood by a person of ordinary skill in the art all language such as “up to,” “at least,” and the like include the number recited and refer to ranges which can be subsequently broken down into subranges or specific values therein as discussed above. Finally, as will be understood by a person of ordinary skill in the art, and as discussed above, a range includes each individual value.

[0049] In understanding the scope of the present disclosure, 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”, “involving”, “having”, and their derivatives.

[0050] The term “consisting” and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other unstated features, elements, components, groups, integers and / or steps.

[0051] The term “consisting essentially of’, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps.

[0052] In embodiments or claims where the term comprising (or the like) is used as the transition phrase, such embodiments can also be envisioned with replacement of the term “comprising” with the terms “consisting of’ or “consisting essentially of.” The methods, kits, systems, and / or devices of the present disclosure can comprise, consist essentially of, or consist of, the components disclosed.

[0053] In embodiments comprising an “additional” or “second” component, the second component as used herein is different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.

[0054] As used herein, “subject” means any animal, preferably a mammal, most preferably a human. The term “mammal” as used herein, encompasses any mammal. Examples of a subject as described herein include but are not limited to fish, birds, reptiles, or mammals, e.g., human, rabbit, cow, pig, sheep, chicken, rat, or mouse.Dielectric Spectroscopy Systems and Devices

[0055] One aspect of the present disclosure relates to a system for collecting or extruding a cell-containing biological material. This system includes an extrusion / collection device comprising an orifice and a chamber operably connected to the orifice. The device also includes a dielectric spectroscopy (DS) device operably connected to the extrusion / collection device. The DS device comprises at least two sets of electrodes, each set comprising a plurality of electrodes. A first set of the at least two sets of electrodes comprises a first material and a first geometrical configuration and a second set of the at least two sets of electrodes comprises a second material and second geometrical configuration. At least one of the first and second materials and the first and second geometrical configurations differ from each other. The DS device is capable of monitoring biologically relevant attributes of the cell -containing biological material as the cellcontaining biological material is extruded from or collected into the extrusion / collection device.

[0056] This aspect of the present disclosure can be combined with any embodiments described for any aspects of the present disclosure.

[0057] The systems and devices described herein (referred to as “the system” or “the device” throughout this disclosure) can use an electrode array that contains different geometries and materials to measure cell concentration in real-time for samples with or without cells and in the presence or absence of flow conditions.

[0058] Using devices and systems described herein, dielectric properties, such as impedance, resistance, reactance, and phase angle, may be detected in non-destructive ways in biological samples or materials containing cell concentrations from about 104to 109cells / mL or any value / range therein, under either static or flow conditions, such as, for example and without limitation, during bioprinting or for cell therapy applications.

[0059] In some embodiments, this is achieved using a dielectric spectroscopy (DS) device, operably connected to an extrusion / collection device, where the DS device is positioned to for the purpose of in-line sensing using multiple electrodes.

[0060] In some embodiments, the multiple electrodes employed in the DS devices vary in orientation, shape, size, distance, and / or material. Additionally, the DS device may comprise two electrode pairs, three electrode pairs, four electrode pairs, five electrode pairs, or any number of electrode pairs suitable for measuring cell concentration in real-time (i.e. the electrode pairs do not display significant cross-talk that obscures the properties measured in a sample).

[0061] In comparison, devices previously reported in the literature for these purposes are limited in their ability to detect such dielectric properties in cell concentrations, because they are only able to detect certain properties in materials containing smaller ranges of cell concentration, z.e., from about 105- 106cells / mL, and such devices have not been reported for use under flow conditions. Since cell and tissue manufacturing often requires producing and processing cell -containing materials in a range of concentration of from about 106to about 109cells / mL, systems and devices of the present disclosure represent a significant improvement over existing devices.

[0062] Extrusion / collection devices are widely known and such devices suitable for inclusion in the systems described herein include, without limitation, devices such as a syringe, cell strainer, valve, tubing, vial, bioreactor, bioprinter, and combinations thereof.

[0063] In some embodiments, the extrusion / collection device is a collection device, and the DS device monitors biologically relevant attributes of the cell containing material as it is collected into the collection device. Suitable collection devices include, without limitation, syringeor syringe-like devices, such as any suitable device capable of collecting biological fluids into a holding container where it would be desirable to monitor biologically relevant attributes of a cellcontaining biological material as it is collected into the collection device. In accordance with this embodiment, the biologically relevant attributes are selected from cell viability, cell count, cell type, cell purity, and combinations thereof.

[0064] In some embodiments, the extrusion / collection device is an extrusion device, and the DS device monitors biologically relevant attributes of the cell-containing biological material as it is extruded from the extrusion device. Suitable extrusion devices include, without limitation, syringe or syringe-like devices, and others, such as those described in U.S. Patent No. 7,939,003; U.S. Patent No. 8,636,938; U.S. Patent No. 8,877,112; and U.S. Patent No. 9,242,031, all of which are hereby incorporated by reference in their entirety. In other words, in the system of the present disclosure, extrusion devices may include a wide array of devices suitable for depositing material to fabricate structures using materials comprising living cells, including material deposition tools described in U.S. Patent No. 7,939,003; U.S. Patent No. 8,636,938; U.S. Patent No. 8,877,112; and U.S. Patent No. 9,242,031, all of which are hereby incorporated by reference in their entirety. A basic structure of a suitable extrusion device is a device comprising some type of collection chamber, or holding chamber where a small to a large amount of material comprising living cells may be stored, and an orifice for extruding the material comprising living cells.

[0065] Dielectric spectroscopy devices are also widely known, and are based on the principle of interaction of an external field with an electric dipole moment of a sample, often expressed by permittivity. DS devices comprise electrodes to create an electrical dipole moment. In the systems and devices described herein, the DS devices comprise at least two sets of electrodes, each comprising a plurality of electrodes. A first set of the at least two sets of electrodes comprises a first material and a first geometrical configuration and a second set of the at least two sets of electrodes comprises a second material and second geometrical configuration.

[0066] Basic principles of dielectric spectroscopy may be observed in a variety of mechanical designs, any of which may be suitable for monitoring biologically relevant attributes of a cell -containing biological material as it is extruded from or collected into an extrusion / collection device. It should be appreciated that certain electrode configurations may confer certain advantages. Electrodes may take various designs, and various designs have been envisioned with the systems and devices described herein, including, for example and without limitation, interdigitated electrodes, an open-ended coaxial probe, coaxial electrodes, parallel plate electrodes, and anti-parallel pin electrodes to name a few. The fundamental requirement of a suitable DSdevice for the system of the present disclosure is that the electric fields generated by the electrodes permeate a cell -containing sample. Beyond that, numerous design configurations may be contemplated. Thus, alternative versions of the DS device may include variations on the electrode geometry / configuration (e.g., parallel plates, coaxial cylinders), the electrode position (e.g., within the syringe barrel, integrated within the needle), or the electrode housing material and configuration (e.g., nylon; cubic, or spherical chambers). Whatever the specific structure, the electrodes should be configured to provide an electrical field sufficient to enable the detection of analytes within a suspending liquid or gel based on changes in impedance.

[0067] In some embodiments, anti-parallel electrode configuration refers to the positional orientation of the electrodes in the extrusion / collection device with the electrodes positioned in opposing directions.

[0068] In some embodiments, an anti-parallel orientation of electrodes can make for easier manufacturing and reduces parasitic effects (e.g., inductances) in the measurements. It should be understood that such an orientation is not critical to device function, and other orientations may also be used (e.g. parallel, interdigitated, coplanar). Whatever orientation the electrodes of a DS device take, they must create an environment in which the electrical field generated by the electrodes can permeate a cell -containing biological material sample.

[0069] In some embodiments, the device can be fabricated by combining multiple electrode types for in-line detection of dielectric properties. The electrodes can vary based on their orientation, shape, size, relative distance to each other, and / or material, and dielectric properties can be collected when the sample being tested / measured is in contact with, or sufficiently affected by, the electric field(s) generated by the electrodes.

[0070] Electrodes may be formed of a variety of conducting materials, including most commonly metals, metal alloys, and graphite materials. In some embodiments, the material used for electrodes of the systems and devices of the present disclosure is a metal selected from gold, platinum, silver, nickel, or tin, or any combination thereof. In some embodiments, the electrode comprises one of these materials or another material and is coated with a metal selected from gold, platinum, silver, nickel, or tin.

[0071] In some embodiments, the material used for electrodes of the systems and devices of the present disclosure is a metal alloy selected from a copper alloy, aluminum alloy, silver alloy, gold alloy, nickel alloy, tin alloy, or any combination thereof. In some embodiments, the electrode comprises one of these metal alloy materials or another material and is coated with a metal alloy selected from a copper alloy, aluminum alloy, silver alloy, gold alloy, nickel alloy, or tin alloy.

[0072] The material of the electrodes of the systems and devices described herein can play a significant role in the ability of the system to detect properties of a material.

[0073] The geometrical configurations of the electrodes of the systems and devices described herein can also play a significant role in the ability of the system to detect properties of a material. In some embodiments, the electrodes comprise a shape that is rod-like. In some embodiments, the electrodes comprise a shape that is flattened, meaning it is rod-like with a flattened side or flattened opposing sides. In some embodiments, the electrodes comprise a shape that is straight. In some embodiments, the electrodes comprise a shape that is curved. In some embodiments, the electrodes comprise a comb-like shape such that the electrodes are interdigitated.

[0074] In some embodiments, pairs of electrodes are identical to each other, meaning the two electrodes that make an electrode pair are constructed of the same material or comprise the same coating and have the same shape or are mirrored images of each other.

[0075] In some embodiments, pairs of electrodes are not identical to each other, meaning the two electrodes that make an electrode pair have different shapes from each other.

[0076] In some embodiments, pairs of electrodes are identical to each other and at least one of the materials and / or geometrical configurations of at least one of the at least two electrode pairs differs from at least another of the at least two electrode pairs. In other words, identical pairs of one set of electrodes in a device / system of the present disclosure differs from another set of identical pairs of electrodes.

[0077] In some embodiments, the system further comprises a system controller comprising a processor and a memory electrically connected to the DS device, where the memory comprises programmed instructions stored thereon and the processor is configured to be capable of executing the stored programmed instructions to: receive one or more of the biologically relevant attributes of the cell-containing biological material from the DS device, and provide instructions to adjust one or more operations of the extrusion / collection device based on the received one or more biologically relevant attributes.

[0078] In some embodiments, operation of the system is controlled by the system controller operably connected to at least the DS device of the system.

[0079] In some embodiments, the extrusion / collection device of the system controls the collection and / or extrusion of a material through an orifice of the extrusion / collection device.

[0080] In some embodiments, impedance measurements, based on data received from the DS device, can be utilized to adjust one or more operations of the extrusion / collection device based on the obtained measurements. For example, the adjustment may adjust the rate or amountof material extruded or collected. In some embodiments, the flow rate of a cell-containing biological material from or into the extrusion / collection device may be varied, the quantity of cells in the cell -containing biological material being extruded from or collected into the extrusion / collection device may be varied, the pressure of the cell -containing biological material as it is extruded from or collected into the extrusion / collection device may be varied, or any combination thereof. Further, the instructions to adjust the one or more operations of the extrusion / collection device may include, for example and without limitation, varying a flow rate of the cell-containing biological material from the extrusion device, varying a quantity of cells in the cell-containing biological material from the extrusion device, varying a pressure of the cellcontaining biological material as the cell -containing material is extruded from the extrusion device, and combinations thereof.

[0081] Biologically relevant attributes monitored by the DS device include, without limitation, cell viability, cell count, cell type, cell purity, and combinations thereof.

[0082] Without being bound by theory, in the present disclosure, it is shown that electrode distance may contribute to signal sensitivity, and electrode shape may impact electric field. Thus, systems and devices of the present disclosure combine two or more different electrode pairs for sample analysis.

[0083] In some embodiments, the device comprises multiple electrode types, which create the ability to increase sensitivity to changes in dielectric properties. With reference to the embodiments illustrated in FIGS. 1 A-1B, one embodiment of device 10 comprises electrode pairs 14 and 16 connected to conducting wire 12, and electrode pairs 14 and 16 are each positioned into a fitting 20, which allows a sample to flow or collect between electrode pairs 14 and 16. An electric current is formed between electrode pairs 14 and 16. When exposed to flow conditions, the sample flows through the passage 8. Device 10 should be capable of undergoing flow rates relevant to the application such as sample collection or extrusion through bioprinting or cell therapy.

[0084] The specific structure of a system and / or device of the present disclosure can be adapted to various applications by shifting the configuration, size, shape, material, or position of the electrodes.

[0085] In some embodiments, one way to create a system and / or device of the present disclosure is to incorporate conductive electrodes into the path of sample transfer. When exposed to flow conditions, the device should be capable of undergoing flow rates relevant to the application such as sample collection or extrusion through bioprinting or cell therapy.

[0086] Thus, a system and / or device described herein is created by integrating conducting electrodes into the transfer mechanism for samples to detect cell concentration with flow rate. As such, it can be adapted to various form factors such as cell strainers, bags, vials, bioreactors, syringes, and bioprinters. Possible non-limiting examples are shown in FIGS. 5B, 6, 17A-17B, and 19, described infra. Additionally, the technology described herein can be applied to various form factors during sample transfer, as depicted in FIG. 18. This includes designs with two pairs of conducting electrodes, each capable of producing an electric field and with a geometry that allows for a sample to flow between both pairs of electrodes.

[0087] In some embodiments, the DS device comprises two pairs of electrodes. With reference again to FIG. 1 A and FIG. IB, illustrated is DS device 10. DS device 10 includes a pair of electrical wires 12 connected to a pair of gold rod electrodes 14 and a pair of electrical wires 12 connected to platinum-coated curved electrodes 16. In FIG. 1 A, fitting 20 is shown with two pairs of electrodes, positioned in an anti-parallel configuration and connected to electrical wires 12. FIG. 1 A is a top view of DS device 10, which shows passage 8, through which biological fluid may flow. As illustrated, gold rod electrodes 14 and platinum curved electrodes 16 are positioned on either side of passage 8, in an anti-parallel configuration, to create an electrical field in which changes in impedance can be measured by each pair of electrodes as biological fluid travels through passage 8. FIG. IB is a bottom view of DS device 10.

[0088] With reference to FIG. 2, illustrated are electrical wires 12 connected to gold rod electrodes 14 before (top) and after (bottom) heat shrink 18 is added.

[0089] With reference to FIG. 3, illustrated is platinum-coated curved electrode 16 adhered to chamber / housing unit 17. As illustrated, chamber / housing unit 17 comprising the platinum- coated curved electrode 16 is held in an upright position using clamps 15.

[0090] In some embodiments, chamber / housing unit 17 comprises openings (not shown) to directly connect electrical wires 12 to chamber / housing unit 17.

[0091] With reference to FIG. 4A, illustrated is a top view of a gold rod electrode device 100 containing gold rod electrodes 14 positioned in fitting 20. The gold rod electrodes 14 are positioned on either side of passage 8. FIG. 4B illustrates that within gold rod electrode device 100 the distance between gold rod electrodes 14 can be varied. This in turn varies the size of passage 8 within fitting 20. Varying the space between electrodes of electrode pairs or between pairs of electrodes can modulate the electrical field and therefore, the sensitivity or ability of a device to monitor biological properties of a material.

[0092] In FIG. 5 A, platinum electrode device 200 is illustrated. Platinum electrode device 200 includes fitting 20 and electrical wires 12 connected to platinum electrodes embedded within fitting 20.

[0093] In some embodiments, systems of the present disclosure have an extrusion / collection device in the form of a syringe or syringe-like device. With reference to FIG. 5B, illustrated is system 300, which is a syringe comprising a Luer type fitting (e.g., a Luer taper fitting) device, which is adapted to include platinum electrode device 200 to monitor biologically relevant attributes of a cell-containing biological material as it enters or exits syringe system 300. System 300 includes extrusion / collection device (syringe) 22 having outer housing 24 with distal end 26 and proximal end 28. Outer housing 24 includes orifice 30 at or near distal end 26.

[0094] With further reference to FIG. 5B, extrusion / collection device (syringe) 22 includes chamber 32, which is defined by outer housing 24. Chamber 32 has an interior volume in which it holds fluid that is meant to exit or enter orifice 30. Thus, orifice 30 is fluidically connected to chamber 32.

[0095] Extrusion / collection device 22 also includes plunger 34. Plunger 34 comprises an elongate rod 36. Elongate rod 36 also includes rod end 38. Rod end 38 is slidably and sealingly engaged with the interior diameter of chamber 32 or, when plunger 34 is in the advanced position, with distal end 26. Plunger 34 is slidably and sealingly insertable into outer housing 24.

[0096] Plunger 34 is axially moveable or slidable within chamber 32 between an advanced position and a retracted position. In the advanced position, plunger rod end 38 is positioned at or near distal end 26. In operation, the advanced position is achieved by pushing or sliding plunger 34 and, in turn, rod end 38 axially toward distal end 26. This can be achieved by a user manually operating plunger 34 or can be achieved by automated mechanical or electromechanical means attached to and operating plunger 34. In the retracted position, rod end 38 is positioned near or toward proximal end 28. In operation, the retracted position is achieved by pulling or sliding plunger 34 and, in turn, rod end 38 axially toward proximal end 28. This can be achieved by a user manually operating the system or can also be achieved by automated mechanical or electromechanical means.

[0097] With further reference to FIG. 5B, system 300 further comprises cannula 40. As described herein, term cannula 40 may include needles or other suitable syringe tips as a type of cannula 40. Fitting 20 may be, for example, a Luer connector. Luer connectors are well known in the art and are a standardized system of small-scale fluid fittings used for creating leak-freeconnections between a male fitting and a mating female part on medical or laboratory instalments, including hypodermic syringe tips and needles or stopcocks and needles.

[0098] With further reference to FIG. 5B, system 300 includes platinum electrode device 200, which includes fitting 20 and electrical wires 12, each of which is connected to an electrode positioned within an in-line channel in fitting 20. Electrical wires 12 are capable of carrying AC current to deliver to electrodes positioned in fitting 20, to create the necessary electrical environment for measuring electrical impedance.

[0099] With further reference to FIG. 5B, fitting 20 is positioned at distal end 26 of extrusion / collection device (syringe) 22. Fitting 20 includes a first end, which connects to distal end 26 via a screw-type fit, and a second end connects to a cannula adapter 42, which connects to cannula 40. Electrical wires 12 lead to electrodes embedded inside of fitting 20 to create an electrical field through which fluid may be monitored. In some embodiments, the internal structure of fitting 20 comprises the electrode structure illustrated in FIGS. 1A-1B (z.e., two pairs of electrodes 14 and 16) or the electrode structure illustrated in FIG. 7 (z.e., three pairs of electrodes 44, 46, and 48).

[0100] In some embodiments, the system of the present disclosure has an extrusion / collection device in the form of a syringe or syringe-like device. Referring to FIG. 6, illustrated is system 400, which is a syringe and Luer type fitting (e.g., a Luer taper fitting) device, which is adapted to include gold rod electrode device 100 to monitor biologically relevant attributes of a cell-containing biological material as it enters or exits syringe 22. System 400 includes extrusion / collection device (syringe) 22 having outer housing 24 with distal end 26 and proximal end 28. Outer housing 24 includes orifice 30 at or near distal end 26.

[0101] With further reference to FIG. 6, extrusion / collection device (syringe) 22 includes chamber 32, which is defined by outer housing 24. Chamber 32 has an interior volume in which it holds fluid that is meant to exit or enter orifice 30. Thus, orifice 30 is fluidically connected to collection chamber 32.

[0102] Extrusion / collection device 22 also includes plunger 34. Plunger 34 comprises an elongate rod 36. Elongate rod 36 also includes rod end 38. Rod end 38 is slidably and sealingly engaged with the interior diameter of chamber 32 or, when plunger 34 is in the advanced position, with distal end 26. Plunger 34 is slidably and sealingly insertable into outer housing 24.

[0103] Plunger 34 is axially moveable or slidable within chamber 32 between an advanced position and a retracted position. In the advanced position, plunger rod end 38 is positioned at or near distal end 26. In operation, the advanced position is achieved by pushing or sliding plunger 34 and, in turn, rod end 38 axially toward distal end 26. This can be achieved bya user manually operating plunger 34 or can be achieved by automated mechanical or electromechanical means attached to and operating plunger 34. In the retracted position, rod end 38 is positioned near or toward proximal end 28. In operation, the retracted position is achieved by pulling or sliding plunger 34 and, in turn, rod end 38 axially toward proximal end 28. This can be achieved by a user manually operating the system or can also be achieved by automated mechanical or electromechanical means.

[0104] With further reference to FIG. 6, system 400 may further comprise cannula 40. As described herein, the term cannula may include needles or other suitable syringe tips as a type of cannula. Fitting 20 may be, for example, a Luer connector.

[0105] With further reference to FIG. 6, system 400 includes a gold rod electrode device 100, which includes fitting 20 and electrical wires 12, each of which is connected to an electrode positioned within an in-line channel in fitting 20. Electrical wires 12 are capable of carrying AC current to deliver to electrodes positioned in fitting 20, to create the necessary electrical environment for measuring electrical impedance.

[0106] With further reference to FIG. 6, fitting 20 is positioned at distal end 26 of extrusion / collection device (syringe) 22. Fitting 20 includes a first end, which connects to distal end 26 via a screw-type fit, and a second end connects to a cannula adapter 42, which connects to cannula 40. Electrical wires 12 lead to electrodes embedded inside of fitting 20 to create an electrical field through which fluid may be monitored. In some embodiments, the internal structure of fitting 20 comprises the electrode structure illustrated in FIG. 1A-1B (z.e., two pairs of electrodes 14 and 16) or the electrode structure illustrated in FIG. 7 (z.e., three pairs of electrodes 44, 46, and 48).

[0107] Referring now to FIG. 7, illustrated is a portion of DS device 500 showing electrode pairs positioned in fitting 20. DS device 500 contains three pairs of electrodes, within fitting 20 and passage 8. The first pair of electrodes are gold electrodes 44 positioned across from each other in fitting 20, the second pair of electrodes are platinum electrodes 46 positioned across from each other in fitting 20, and the third pair of electrodes are silver electrodes 48 positioned across from each other in fitting 20. Thus, in this DS device 500 illustrated in FIG. 7, pairs of gold (Au) electrodes 44, platinum (Pt) electrodes 46, and silver (Ag) electrodes 48 of varying distances and shapes are utilized.

[0108] In reference now to FIG. 14, illustrated is gold rod electrode device 100, which has fitting 20, with embedded gold rod electrodes 14 connected to electrical wires 12. The illustrated gold rod electrode device 100 shows passage 8, through which biological fluid may flow. Fitting20 and electrical wires 12 are surrounded by epoxy 9. Epoxy 9 is used to seal the gold rod electrode device 100 and prevent leakage.

[0109] In another embodiment, the system of the present disclsoure has an extrusion / collection device in the form of a syringe or syringe-like device. Referring now to FIG. 19, illustrated is system 600, which is a syringe and Luer type fitting (e.g., a Luer taper fitting) device, which is adapted to include gold rod electrode device 100 and platinum electrode device 200 in series to monitor biologically relevant attributes of a cell-containing biological material as it enters or exits the syringe. System 600 includes extrusion / collection device (syringe) 22 having outer housing 24 with distal end 26 and proximal end 28.

[0110] With further reference to FIG. 19, extrusion / collection device (syringe) 22 includes chamber 32, which is defined by outer housing 24. Chamber 32 has an interior volume in which it holds fluid.[oni] Extrusion / collection device 22 also includes plunger 34. Plunger 34 comprises an elongate rod 36. Elongate rod 36 also includes rod end 38. Rod end 38 is slidably and sealingly engaged with the interior diameter of collection chamber 32 or, when plunger 34 is in the advanced position, with distal end 26. Plunger 34 is slidably and sealingly insertable into outer housing 24.

[0112] Plunger 34 is axially moveable or slidable within chamber 32 between an advanced position and a retracted position. In the advanced position, plunger rod end 38 is positioned at or near distal end 26. In operation, the advanced position is achieved by pushing or sliding plunger 34 and, in turn, rod end 38 axially toward distal end 26. This can be achieved by a user manually operating plunger 34 or can be achieved by automated mechanical or electromechanical means attached to and operating plunger 34. In the retracted position, rod end 38 is positioned near or toward proximal end 28. In operation, the retracted position is achieved by pulling or sliding plunger 34 and, in turn, rod end 38 axially toward proximal end 28. This can be achieved by a user manually operating the system or can also be achieved by automated mechanical or electromechanical means.

[0113] With reference to FIG. 19, system 600 may further comprise cannula 40. As described herein, the term cannula may include needles or other suitable syringe tips as a type of cannula. Fitting 20 may be, for example, a Luer connector.

[0114] With further reference to FIG. 19, system 600 includes a gold rod electrode device 100, which includes fitting 20 and electrical wires 12, each of which is connected to an electrode positioned within an in-line channel in fitting 20. The system 600 also includes a platinum electrode device 200, which includes fitting 20 and electrical wires 12, each of which isconnected to an electrode positioned within an in-line channel in fitting 20. Electrical wires 12 are capable of carrying AC current to deliver to electrodes positioned in fitting 20, to create the necessary electrical environment for measuring electrical impedance. In some embodiments, the internal structure of one or both of fittings 20 comprises, independently, the electrode structure illustrated in FIGS. 4A-4B (z.e., one pair of electrodes 14), FIG. 14 (z.e., one pair of electrodes 14), FIGS. 1 A-1B (z.e., two pairs of electrodes 14 and 16), or the electrode structure illustrated in FIG. 7 (z.e., three pairs of electrodes 44, 46, and 48). Thus, in some embodiments, the electrode pairs are in the same device, while in some embodiments, the electrode pairs are in separate devices in series.

[0115] Another aspect of the present disclosure relates to a system for collecting or extruding a cell-containing biological material. The system includes a syringe device comprising a housing having opposed distal and proximal ends, where the distal end comprises an orifice. The syringe device also includes a fitting connected at the orifice for interchangeably connecting a needle, a plunger axially movable within the housing between an advanced position near the distal end and a retracted position near the proximal end, and an elongate rod being connected to the plunger to move the plunger axially between the advanced and retracted positions and extending through the proximal end of the housing. The system also includes a dielectric spectroscopy (DS) device operably connected to the syringe device at the fitting. The DS device comprises a first set of electrodes and a second set of electrodes, the first set of electrodes comprises a first material and a first geometrical configuration, and the second set electrodes comprises a second material and a second geometrical configuration. At least one of the first and second materials and the first and second geometrical configurations differ from each other. The electrodes create an electrical field for detecting impedance of a cell -containing biological material as the cell -containing biological material is extruded from or collected into the syringe device.

[0116] This aspect of the present disclosure can be combined with any embodiments described for any aspects of the present disclosure.

[0117] In some embodiments, the fitting is a Luer taper fitting. In some embodiments, the fitting is customized from a female luer lock. In some embodiments, the fitting is a custom- made housing unit. In accordance with this embodiment, any housing-unit that maintains the electrodes at a fixed distance is suitable for use in the devices and systems disclosed herein. The custom-made housing unit can be fitted to a syringe by adding luer locks to either end.

[0118] The disclosed systems and devices described herein are highly adaptable. Accordingly, the fitting may be substituted for an appropriate adapter dependent on the form factor the system and devices are adapted to.

[0119] In some embodiments, the DS devices described supra may be coated with an epoxy. The epoxy coating is used to seal the DS device to prevent leakage. In accordance with this embodiment, alternative adhesives and resins that prevent leakage may be utilized to coat the DS devices of the present disclosure.

[0120] In some embodiments, devices described herein may be designed to attach to a syringe for real-time, in-line data collection during sample extrusion or collection. The syringe device can then detect dielectric properties of a sample to be tested / measured and of cells within the sample, enabling the device to detect cellular properties such as cell concentration. Notably, this technology may have additional uses, for example using dielectric spectroscopy to detect cell type in cell-laden alginate bioink, distinguishing between the dielectric properties of various cell types in bioprinted constructs post-fabrication, detecting cell viability in samples without flow rate, or other suitable uses.

[0121] A further aspect of the present disclosure is directed to a dielectric spectroscopy (DS) device connectable to an extrusion / collection device, where the DS device comprises antiparallel electrodes configured to create an electrical field around a fluid passageway for detecting impedance of a cell -containing biological material as it moves through the passageway.

[0122] In some embodiments, this aspect of the present disclosure relates to a system comprising an in-line DS device that is capable of monitoring biologically relevant attributes of a cell -containing biological material as it is extruded from an extrusion device (e.g., in a bioprinting context, or in administering a cell-containing solution to a subject as part of cell therapy) or as it is collected into a collection device (e.g., from a subject or an in vitro setting). Monitoring is accomplished through the DS device operably coupled to an extrusion / collection device.

[0123] A further aspect of the present disclosure relates to a dielectric spectroscopy (DS) device connectable to an extrusion / collection device, where the DS device comprises a first and second set of anti-parallel electrodes. The first set of anti-parallel electrodes comprises a first material and a first geometrical configuration and the second set of anti-parallel electrodes comprising a second material and a second geometrical configuration. At least one of the first and second material and the first and second geometrical configuration differ from each other. The first and second set of anti-parallel electrodes are configured to create an electrical field around a fluid passageway for detecting impedance of a cell -containing biological material as the cell -containing biological material moves through the fluid passageway.

[0124] This aspect of the present disclosure can be combined with any embodiments described for any aspects of the present disclosure.

[0125] Another aspect of the present disclosure relates to an extrusion / collection device comprising an orifice and a chamber operably connected to the orifice. The extrusion / collection device is configured to be operably connected to a dielectric spectroscopy (DS) device comprising a first and a second set of electrodes, the first set of electrodes comprising a first material and a first geometrical configuration, the second set of electrodes comprising a second material and a second geometrical configuration. At least one of the first and second material and the first and second geometrical configuration differ from each other. The DS device is configured to monitor biologically relevant attributes of a cell-containing biological material as the cell-containing biological material is extruded from or collected into the extrusion / collection device.

[0126] This aspect of the present disclosure can be combined with any embodiments described for any aspects of the present disclosure.

[0127] A further aspect of the present disclosure relates to a system for measuring a parameter of a biological material. The system includes a material receiving device configured to contact the biological material, the material receiving device comprising an orifice and a chamber operably connected to the orifice, the chamber being configured to receive the biological material therein. The system also includes a dielectric spectroscopy (DS) device comprising an electrode operably connected to the material receiving device. The DS device is configured to monitor a parameter of the biological material when the biological material is received in the material receiving device.

[0128] This aspect of the present disclosure can be combined with any embodiments described for any aspects of the present disclosure.Methods

[0129] Examples of applications of the disclosed systems and devices can include changes in form factors such as: bioprinting or syringe pump (syringe to printing bed, product package to syringe), cell therapy (patient to syringe, syringe to patient, syringe to cell strainer, syringe to bioreactor, bioreactor to bioreactor), cell manufacturing (syringe to flask, syringe to bioreactor, syringe to cell strainer, flask to bioreactor, flask to cell strainer, bioreactor to bioreactor), industrial or lab-scale cell processing (patient to syringe, syringe to patient, syringe to cell strainer, syringe to bioreactor, bioreactor to bioreactor, syringe to flask, flask to bioreactor, flask to cell strainer), patient to infusion station, cell isolation, etc.

[0130] Thus, another aspect of the present disclosure relates to a method of depositing a cell -containing biological material. This method involves providing a system as described herein, where the extrusion / collection device is an extrusion device, extruding from the extrusion device the cell-containing biological material, and monitoring biologically relevant attributes of the cellcontaining biological material as it is extruded from the device.

[0131] In some embodiments, extruding comprises printing a living, three-dimensional tissue.

[0132] In some embodiments, the cell-containing biological material comprises a cell therapy composition. In some embodiments, extruding comprises the cell-containing biological material into a subject to administer cell therapy treatment. In some embodiments, the biologically relevant attributes are selected from the group consisting of cell viability, cell count, cell type, cell purity, and combinations thereof.

[0133] In some embodiments, the method of this aspect further involves adjusting the extrusion of the cell -containing biological material based on the monitored biologically relevant attributes of the cell-containing biological material.

[0134] A further aspect of the present disclosure relates to a method of collecting a cellcontaining biological material. This method involves providing a system as described herein, where the extrusion / collection device is a collection device, collecting into the collection device the cell-containing biological material, and monitoring biologically relevant attributes of the cellcontaining biological material as it is collected into the device. In some embodiments, the biologically relevant attributes are selected from the group consisting of cell viability, cell count, cell type, cell purity, and combinations thereof.

[0135] In some embodiments, the collection of the cell-containing biological material is adjusted based on the monitored biologically relevant attributes of the cell -containing biological material. The technology addressed in this disclosure non-destructively assesses cell concentration across the 104- 109cells / mL range in real-time using dielectric spectroscopy.Through experiments, it was also determined that relevant flow rates within the 10'1- 103mL / min range can be compatible with the disclosed technology. Previously created devices (e.g., U.S.Patent No. 12,019,040, which is hereby incorporated by reference in its entirety) can be used to measure cell concentration in real-time for bioprinting. Such devices can be used with syringes and related materials. The technology disclosed herein is not limited only to use with syringes and related materials and offers an adaptable design that can be incorporated into various systems during sample transfer.

[0136] Measuring cell concentration during sample transfer relies on the consistency of the signal reading for accurate data collection. In FIG. 15, an exemplary embodiment of the device can measure real-time changes in cell concentration over 420 seconds (7 minutes). Meanwhile, FIGs. 16A and 16B show that embodiments of the device can produce consistent results for homogenous samples with set cell concentration at varied flow rates. This indicated that flow rate did not impact measurement of bulk cell concentration.

[0137] In some embodiments, the systems and devices can be used in various form factors such as, but not limited to: bioprinting or syringe pump (syringe to printing bed, product package to syringe), cell therapy (patient to syringe, syringe to patient, syringe to cell strainer, syringe to bioreactor, bioreactor to bioreactor), cell manufacturing (syringe to flask, syringe to bioreactor, syringe to cell strainer, flask to bioreactor, flask to cell strainer, bioreactor to bioreactor), industrial or lab-scale cell processing (patient to syringe, syringe to patient, syringe to cell strainer, syringe to bioreactor, bioreactor to bioreactor, syringe to flask, flask to bioreactor, flask to cell strainer, cell strainer to tube), patient to infusion station, cell isolation, etc. In addition to measuring cell concentration, such data can be used to implement feedback control of transfer to achieve desired rates of cell deposition or cell transfer. Such control would obviate the need for additional measurements as well as enable the fabrication of more complex tissues via bioprinting.

[0138] The systems, devices, and methods described herein can be applied to a wide variety of form factors for in-line, real-time measurement of cellular properties during sample transfer. FIGS. 17A-17B show two examples: bioprinter to bioprinter bed (FIG. 17A) and syringe pump to tube (FIG. 17B).

[0139] The systems and devices of the present disclosure may be used in methods of fabricating an article and methods of fabricating a living three-dimensional structure, where that method involves, e.g., extruding cell -containing biological material from an extrusion device. General methods of fabricating a living three-dimensional structure are described herein and may involve the system and methods also described in U.S. Patent No. 7,939,003; U.S. Patent No. 8,636,938; U.S. Patent No. 8,877,112; and U.S. Patent No. 9,242,031, all of which are hereby incorporated by reference in their entirety. The advantage of the system of the present disclosure is that it has the ability to monitor biologically relevant attributes of a cell-containing biological material as it is extruded from the extrusion device, as well as adjusting the rate / amount of extruded material based on the monitored biologically relevant attributes.

[0140] In some embodiments, methods described herein further comprises adjusting the extrusion of the cell -containing biological material based on the monitored biologically relevant attributes of the cell-containing biological material.

[0141] Cell-containing biological materials that may be used with the system and methods of the present disclosure include, without limitation, any material capable of being deposited from a material deposition tool onto a substrate. The type of material will depend on the type of material deposition tool (or extrusion device) being employed. For example, when a syringe-like device is employed, suitable materials include, without limitation, virtually any liquid, slurry, or gel.

[0142] In some embodiments, the cell-containing biological material is a hydrogel having seeded cells. Suitable hydrogels include, without limitation, alginate, agarose, collagen, chitosan, fibrin, hyaluronic acid, carrageenan, polyethylene oxide, polypropylene oxide, polyethylene oxide- co-polypropylene oxide, hydroxypropyl methyl cellulose, polypropylene fumarate-co-ethylene glycol), poly(ethylene glycol)-co-poly(lactic acid), poly(vinyl alcohol), KDL12 oligopeptides, and poly(n-isopropyl acrylamide).

[0143] Hydrogels may have a controlled rate of crosslinking through the adjustment of environmental variables including, but not limited to, temperature, pH, ionic strength, heat, light, or the addition of chemical crosslinking agents such as calcium, magnesium, barium, chondroitin, sulfate, and thrombin. A cross-linking compound may be provided in a weight ratio of hydrogel to cross-linking compound of about 1 :100 to 100: 1, respectively. In some embodiments, the weight ratio of cross-linking compound to hydrogel is about 1 :5.3. In some embodiments, the crosslinking compound is calcium sulfate.

[0144] In some embodiments, cells in a hydrogel are of a single cell type. Suitable cell types include, without limitation, all mammalian or plant cells. Specific cell types may include, without limitation, chondrocytes, osteoblasts, osteoclasts, osteocytes, fibroblasts, hepatocytes, skeletal myoblasts, cardiac myocytes, epithelial cells, endothelial cells, keratinocytes, neurons, Schwann cells, oligodendrocytes, astrocytes, pneumocytes, adipocytes, smooth muscle cells, T cells, B cells, marrow-derived stem cells, hematopeotic stem cells, osteoprogenitor cells, neural stem cells, and embryonic stem cells. In some embodiments, cells in the hydrogel may be of more than one cell type.

[0145] Dispensing material from an extrusion device may be carried out under sterile conditions including, for example, in a hermetically sealed envelope or container.

[0146] In some embodiments, systems, devices, and / or methods of the present disclosure may be used in administering to a subject a cell-containing biological material for which monitoring biologically relevant attributes of the cell-containing biological material is desirable.For example, in cell therapy, cell transplantation, cytotherapy, and other therapies that involve the administration of living cells to a subject, it would be desirable to monitor the attributes of the cellcontaining biological material as it is being administered from an extrusion device (e.g., a syringe). Using a system and / or device of the present disclosure, one could monitor the quantity, quality, and even type of cells being injected, grafted, or implanted into a subject to affect a medicinal effect. Further, the rate / amount of deposited material can be adjusted based on the monitoring.

[0147] Thus, a further aspect relates to a method of collecting a cell-containing biological material. This method involves providing a system or device as described herein, collecting into the device a cell-containing biological material, and monitoring biologically relevant attributes of the cell-containing biological material as it is collected into the device.

[0148] In some embodiments, the device can be used for real-time measurement of cell concentration for samples during transport. Because data is collected in real-time, the sample can be monitored over time with flow rate for real-time feedback during fabrication. Combining multiple electrodes into a single device or series of devices can greatly increase data generated for the sample, leading to a future need for machine learning or automated data analysis processes.

[0149] In some embodiments, the present disclosure relates to a method of collecting a cell -containing biological material. This method involves providing the system according to the present disclosure, collecting into the collection device the cell-containing biological material, and monitoring biologically relevant attributes of the cell-containing biological material as the cell -containing biological material is collected into the device. In accordance with these embodiments, the biologically relevant attributes are selected from cell viability, cell count, cell type, cell purity, and combinations thereof. Further, the method can comprise adjusting the collection of the cell-containing biological material based on the monitored biologically relevant attributes of the cell-containing biological material.

[0150] Meanwhile, cell therapy treatments can be drastically improved with real-time monitoring of cell count. For example, current techniques for harvesting stem cells from patient bone marrow rely on guesswork and overharvesting to obtain the required minimum cells for treatment. These inaccuracies arise from the inability to assess cell concentration until after collection. Notably, withdrawal of too much marrow is both painful and potentially harmful to patients. The disclosed device can give immediate information, providing an intuitive understanding of how much sample is needed per patient.

[0151] In some embodiments, cell -containing biological material comprises a cell therapy composition. In some embodiments, the extruding comprises extruding the cellcontaining biological material into a subject to administer the cell therapy treatment.

[0152] Collection using the system and methods of the present disclosure may occur in a laboratory or in vitro setting where monitoring of fluids collected from containers or other laboratory equipment is desirable. In some embodiments, the system and methods of the present disclosure may involve collecting cell-containing biological material directly from subjects. Regardless, if it is important or desirable to monitor biologically relevant attributes of the cellcontaining biological material being collected, the system and methods of the present disclosure, which incorporate a DS device, is able to provide information relating to the biologically relevant attributes of the cell-containing biological material. In certain embodiments, the system also adjusts the collection based on the obtained information.

[0153] The systems and methods of the present disclosure involve monitoring biologically relevant attributes of a cell-containing biological material being extruded from or collected into the extrusion / collection device. As used herein, biologically relevant attributes of a cell -containing biological material, which may be monitored with the DS device described herein, include, without limitation cell viability, cell count, cell type, cell purity, and combinations thereof. Cell viability is a measure of living or intact cells in the biological material (ie., un-lysed cells); cell count includes a measure of the number of cells in a biological material, the total number of cells extruded out of or into a collection device, or the rate of cell deposition or intake over a period of time; cell type is a measure of cell type, e.g., bacterial cells versus eukaryotic or mammalian cells; and cell purity is a measure of the purity of a cell-containing solution by determining the presence or absence of other contaminants in a solution, or different types of cells in a solution, e.g., if it is desirable to have a solution with a single cell type. For example, the system and methods of the present disclosure may involve 3D bioprinter quality monitoring, injectable drug delivery quality assurance, or chemical or biological contamination detection in laboratory or research settings. Noninvasive, nontoxic, and nondestructive real-time monitoring of biologically active materials is a largely unmet challenge in biomedical engineering. Nondestructive means of assessment of critical quality attributes of these materials is especially of interest; the time and / or cost associated with generating biological samples such as tissue constructs is at odds with the currently widespread destructive biochemical analysis of such materials and hinders research in this area. Further, regulatory concerns exist in gathering data from sacrificial samples as a proxy for measurements taken directly from patient-bound materials. To address these concerns, the monitoring approach described herein is nondestructive to biological materials given sufficient calibration and is integrated within the material delivery or collection environment, e.g., a syringe or syringe-compatible channel.EXAMPLES

[0154] The examples below are intended to exemplify the practice of embodiments of the disclosure but are by no means intended to limit the scope thereof.Materials and Methods

[0155] One exemplary device incorporates electrodes into a chamber that attaches to a syringe. As the sample is collected or extruded, the sample is in contact with the electric field(s), and the dielectric properties of impedance, resistance, reactance, and phase angle are detected in real-time. Because the electrodes may vary greatly in fabrication, shape, and material, an example of electrode formation is shown in FIG. 1 A-1B. Considering the two electrode examples shown in FIG. 1A-1B, the fabrication of rod and curved electrodes will be described.Rod Electrodes

[0156] Exemplary gold rod-shaped electrodes were experimentally determined to be sensitive to cell concentrations from 106-l 09cells / mL. These electrodes were crimped onto insulated copper wires, sealed with heat-shrink, and added to the device chamber, as depicted in FIG. 2.Curved Electrodes

[0157] Exemplary platinum-coated curved electrodes were determined to be sensitive to cell concentrations from 104- 106cells / mL. These electrodes can be manufactured by sputtercoating platinum. Copper wires can be threaded through the device chamber and connected to the platinum electrodes using silver epoxy or another suitable material, as shown in FIG. 3. Both halves of the device chamber can be combined with male and female luer locks, for example by using epoxy.Example 1 - Multi-Electrode Device

[0158] Experimental data has been generated for prototypes that utilize rod-shaped gold electrodes and platinum-coated curved electrodes (FIGs. 1-3) in two separate prototypes.Illustrations of each are shown in FIGs. 4-6. A multi-electrode device can be fabricated by combining the two electrode designs into a single chamber, as depicted in FIG. 1. All versions of this device are compatible with multiple syringes and syringe tips, as depicted in FIG. 6.

[0159] The performance of each prototype (FIGs. 4-6) was evaluated and compared by assessing the dielectric property of impedance at low (0-4.5 x 106cells / mL) and high (20 x 106- 70 x 106cells / mL) cell concentrations using neonatal bovine-derived primary chondrocytes suspended in phosphate buffered saline in FIG. 8. Device sensitivity, a measure of the change inimpedance or phase angle as cell concentration changes, was determined for each electrode type and shows that each electrode type is best suited to detect cell concentration at different ranges (FIG. 9).

[0160] Cell concentration of primary bovine-derived chondrocytes was also assessed from 1 x 106-125 x 106cells / mL with n=3 using only the gold rod-shaped electrode prototype, shown in FIG. 10. Control measurements of PBS were collected to remove the background and isolate the impedance (Z) of the cells (Zceii, Equation 1). To assess repeatability, this study was conducted 5 times using primary chondrocytes from different neonatal joints on different days. In addition, phase angle was assessed in FIG. 11 by removing the PBS measurement from the cell sample (0ceii). Lastly, device sensitivity was determined to vary between impedance and phase angle but was highest at low frequencies below 100 kHz (FIGs. 12-13).Example 2 - Multi-Electrode Device in Series

[0161] Referring to FIG. 19, a multi-electrode prototype utilizing gold rod electrodes and curved platinum electrodes is depicted. The device was created by combining the individual smart syringes in series (FIG. 19). Primary chondrocytes suspended in phosphate buffered saline (PBS) were assessed by ranging the cell concentration from 0.6 - 112 x 106cells / mL. Control measurements of PBS were collected to remove the background and isolate the impedance (Z) of the cells (Zceii, Equation 1). Measurements were normalized to the PBS readings to obtain the fractional change (Equation 2). It was found that from 0.5 kHz to 60 kHz, both electrode types displayed frequency-dependent separation between the cell concentrations (FIG. 20A-20B, Equation 2). Frequencies from 60-1,000 kHz were polluted with electrode crosstalk and were thus unusable. To further assess the capability and resolution of each electrode, impedance measurements from specific frequencies were graphed in FIG. 21 and FIG. 22. ceii ZsampieZpp (Equation 1) (Equation 2)

[0162] The curved platinum electrodes showed the greatest sensitivity to cell concentration at 0.1 kHz and 1 kHz, which is seen in the changes in Z fractional change from 0.6 x 106- 56 x 106cells / mL (FIG. 21A and FIG. 21B). In contrast, the gold rod electrodes were better suited to 10 kHz as denoted by the increase in Z fractional change from 28.5-112 x 106cells / mL (FIG. 21C). At 100kHz, both electrodes experienced cross-talk and failed to show sufficient trends (FIG. 21D).

[0163] The findings show that a multi -el ectrode device can provide heighted sensitivity to a greater range of cell concentrations than would be assessable to either electrode alone. In addition, the electrodes may show improved resolution at different frequencies.

[0164] A piecewise linear regression was used to mark trends in behavior (FIG. 22). Gold rod electrodes display decreasing impedance with increasing cell concentration at 0.1 kHz and 1 kHz, the opposite behavior from the curved platinum electrodes (FIG. 22A and FIG. 22B). At 10 kHz, both electrodes display increasing impedance with cell concentration, however, impedance is lower at 10 kHz compared to 0.1 kHz or 1 kHz, marking a decrease in sensitivity (FIG. 22C). Lastly, data collection was impacted by electrode cross-talk at 100 kHz (FIG. 22D).

[0165] Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the claims which follow.

Claims

WHAT IS CLAIMED IS:

1. A system for collecting or extruding a cell -containing biological material, said system comprising: an extrusion / collection device comprising: an orifice; and a chamber operably connected to the orifice; and a dielectric spectroscopy (DS) device operably connected to the extrusion / collection device, wherein the DS device comprises at least two sets of electrodes, each set comprising a plurality of electrodes, wherein a first set of the at least two sets of electrodes comprises a first material and a first geometrical configuration and a second set of the at least two sets of electrodes comprises a second material and a second geometrical configuration, wherein at least one of the first and second material and the first and second geometrical configuration differ from each other, and wherein the DS device is capable of monitoring biologically relevant attributes of the cell -containing biological material as the cell -containing biological material is extruded from or collected into the extrusion / collection device.

2. The system according to claim 1, wherein the extrusion / collection device comprises a syringe, cell strainer, valve, tubing, vial, bioreactor, bioprinter, and combinations thereof.

3. The system according to claim 1, wherein at least one of the first and second material comprises a metal selected from gold, platinum, or a combination thereof.

4. The system according to claim 3, wherein at least one of the first and second material is coated with a metal selected from gold or platinum.

5. The system according to claim 1, wherein at least one of the electrodes comprises a shape that is rod-like, flattened, or curved.

6. The system according to claim 1 further comprising:a system controller comprising a processor and a memory electrically connected to the DS device, wherein the memory comprises programmed instructions stored thereon and the processor is configured to be capable of executing the programmed instructions to: receive one or more of the biologically relevant attributes of the cellcontaining biological material from the DS device; and provide instructions to adjust one or more operations of the extrusion / collection device based on the received one or more biologically relevant attributes.

7. The system according to claim 6, wherein the instructions to adjust the one or more operations of the extrusion / collection device are selected from the group consisting of varying a flow rate of the cell -containing biological material from the extrusion device, varying a quantity of cells in the cell-containing biological material from the extrusion device, varying a pressure of the cell -containing biological material as the cell -containing material is extruded from the extrusion device, and combinations thereof.

8. The system according to claim 1, wherein the extrusion / collection device is an extrusion device, and said DS device monitors biologically relevant attributes of the cellcontaining biological material as it is extruded from the extrusion device.

9. The system according to claim 1, wherein the extrusion / collection device is a collection device, and said DS device monitors biologically relevant attributes of the cellcontaining material as it is collected into the collection device.

10. The system according to claim 1, wherein the biologically relevant attributes are selected from the group consisting of cell viability, cell count, cell type, cell purity, and combinations thereof.

11. A method of depositing a cell-containing biological material, said method comprising: providing the system according to claim 8; extruding from the extrusion device the cell -containing biological material; and monitoring biologically relevant attributes of the cell-containing biological material as it is extruded from the device.

12. The method according to claim 11, wherein said extruding comprises printing a living, three-dimensional tissue.

13. The method according to claim 11, wherein the cell-containing biological material comprises a cell therapy composition.

14. The method according to claim 13, wherein said extruding comprises extruding the cell -containing biological material into a subject to administer the cell therapy treatment.

15. The method according to claim 13, wherein the biologically relevant attributes are selected from the group consisting of cell viability, cell count, cell type, cell purity, and combinations thereof.

16. The method according to claim 14 further comprising: adjusting the extrusion of the cell -containing biological material based on the monitored biologically relevant attributes of the cell-containing biological material.

17. A method of collecting a cell -containing biological material, said method comprising: providing the system according to claim 9; collecting into the collection device the cell-containing biological material; and monitoring biologically relevant attributes of the cell-containing biological material as the cell-containing biological material is collected into the device.

18. The method according to claim 17, wherein the biologically relevant attributes are selected from the group consisting of cell viability, cell count, cell type, cell purity, and combinations thereof.

19. The method according to claim 18 further comprising: adjusting the collection of the cell -containing biological material based on the monitored biologically relevant attributes of the cell-containing biological material.

20. A system for collecting or extruding a cell -containing biological material, said system comprising: a syringe device comprising: a housing having opposed distal and proximal ends, wherein the distal end comprises an orifice; a fitting connected at the orifice for interchangeably connecting a needle; a plunger axially movable within the housing between an advanced position near the distal end and a retracted position near the proximal end; an elongate rod being connected to the plunger to move the plunger axially between the advanced and retracted positions, and extending through the proximal end of said housing; and a dielectric spectroscopy (DS) device operably connected to the syringe device at the fitting, wherein the DS device comprises a first set of electrodes and a second set of electrodes, the first set of electrodes comprising a first material and a first geometrical configuration, and the second set electrodes comprises a second material and a second geometrical configuration, wherein at least one of the first and second materials and the first and second geometrical configurations differ from each other, and wherein the first and second set of electrodes are configured to create an electrical field for detecting impedance of a cell-containing biological material as the cell -containing biological material is extruded from or collected into the syringe device.

21. The system according to claim 20, wherein the fitting is a Luer taper fitting.

22. The system according to claim 20 further comprising: a system controller comprising a processor and a memory electrically connected to the DS device, the memory comprising programmed instructions stored thereon and the processor being configured to be capable of executing the stored programmed instructions to: receive one or more of the biologically relevant attributes of the cellcontaining biological material from the DS device; and provide instructions to adjust one or more operations of the syringe device based on the received one or more biologically relevant attributes.

23. The system according to claim 22, wherein the instructions to adjust one or more operations of the syringe device is selected from the group consisting of varying a flow rate of the cell-containing biological material from the extrusion device, varying a quantity of cells in the cell-containing biological material from the extrusion device, varying a pressure of the cell -containing biological material as the cell -containing biological material is extruded from the extrusion device, and combinations thereof.

24. A dielectric spectroscopy (DS) device connectable to an extrusion / collection device, wherein the DS device comprises a first and second set of antiparallel electrodes, the first set of anti-parallel electrodes comprising a first material and a first geometrical configuration and the second set of anti-parallel electrodes comprising a second material and a second geometrical configuration, wherein at least one of the first and second material and the first and second geometrical configuration differ from each other, and wherein the first and second set of anti-parallel electrodes are configured to create an electrical field around a fluid passageway for detecting impedance of a cell-containing biological material as the cell -containing biological material moves through the fluid passageway.

25. The DS device according to claim 24 further comprising: a system controller comprising a processor and a memory electrically connected to the DS device, the memory comprising programmed instructions stored thereon and the processor being configured to be capable of executing the stored programmed instructions to receive one or more biologically relevant attributes of the cell-containing biological material from the DS device.

26. The DS device according to claim 25, wherein the biologically relevant attributes are selected from the group consisting of cell viability, cell count, cell type, cell purity, and combinations thereof.

27. An extrusion / collection device comprising: an orifice; and a chamber operably connected to the orifice,wherein the extrusion / collection device is configured to be operably connected to a dielectric spectroscopy (DS) device comprising a first and a second set of electrodes, the first set of electrodes comprising a first material and a first geometrical configuration, the second set of electrodes comprising a second material and a second geometrical configuration, wherein at least one of the first and second material and the first and second geometrical configuration differ from each other, and the DS device being configured to monitor biologically relevant attributes of a cell-containing biological material as the cell -containing biological material is extruded from or collected into the extrusion / collection device.

28. A system for measuring a parameter of a biological material, said system comprising: a material receiving device configured to contact the biological material, the material receiving device comprising an orifice and a chamber operably connected to the orifice, the chamber being configured to receive the biological material therein; and a dielectric spectroscopy (DS) device comprising an electrode and being operably connected to the material receiving device, wherein the DS device is configured to monitor a parameter of the biological material when the biological material is received in the material receiving device.

29. The system of claim 28, wherein the parameter monitored by the DS device is a biologically relevant attribute of a cell -containing biological material.

30. The system of claim 28 or 29, wherein the material receiving device comprises at least two electrodes comprising a material and geometrical configuration.

31. The system of claim 30, wherein at least one of the material and geometrical configuration of at least one of the at least two electrodes differs from at least another of the at least two electrodes.

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