Enhanced cell and biologics production using interconnecting porous hydrogel blocks

Abstract

Described herein are modular cell culture systems, methods, and interconnecting porous hydrogel blocks. A block may comprise a 3D continuous polymeric matrix with a network of microporous cavities, and may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal. A system may comprise at least one block so configured. A method may comprise seeding at least one block so configured with at least one cell type.

Description

[0001] ENHANCED CELL AND BIOLOGICS PRODUCTION USING INTERCONNECTING POROUS HYDROGEL BLOCKS

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] This application claims the benefit of and priority to U.S. Provisional Application Serial No. 63 / 666,413, filed July 1, 2024, which is hereby incorporated by reference in its entirety for all that it contains (including all references therein) for all purposes as if restated and set forth fully herein to the maximum extent allowable by law.

[0004] TECHNICAL FIELD

[0005] The subject matter disclosed herein is generally directed to the field of cell and biologic production, and the use of modular tissue-mimetic platforms such as interconnecting porous hydrogel blocks (IPHBs), e.g. to enhance and streamline the production of mammalian cells and / or biologies. IPHBs and embodiments herein may provide scalable and / or physiologically relevant environments), such as for co-culture of multiple cell types, e.g. enabling formation of organized tissue structure(s) and / or functional organ components). This disclosure addresses challenges associated with traditional cell culture methods and current bioreactor technologies, offering transformative solutions that improve cell viability, functionality, and / or yield.

[0006] BACKGROUND

[0007] The advancement of drug discovery and biotherapeutic development relies on the ability to utilize and test representative models that mimic human physiology. Traditional in vitro technologies for cultivating cells, such as plastic dishes, plates, or flasks, fall short in replicating the complexity and functionality of human tissues and organs. These conventional methods often lead to inaccurate or false results, as cells do not behave the same way in a two- dimensional (2D) environment as they do in their native three-dimensional (3D) context. Additionally, these technologies lack the adaptability, flexibility, and versatility required for mimicking the diverse microenvironments found in human tissues.

[0008] Scaling the production of cells and biologies presents numerous challenges, particularly within current Good Manufacturing Practice (cGMP) environments. These challenges span technical, operational, regulatory, and economic aspects. Technical challenges include maintaining cell quality and consistency, optimizing culture conditions, and scalability of equipment. Ensuring uniformity and consistency in cell cultures is difficult, as variations in cell phenotype and functionality may impact the quality of the final product. Efficient nutrient supply and waste removal are important for maintaining cell viability and productivity in large-scale cultures. Traditional bioreactors have limitations in scaling without compromising cell health and productivity.

[0009] Operational challenges include process standardization and facility design and infrastructure, and achieving reproducibility and consistency in large-scale production processes. Adequate space and controlled environments are important for scaling production.

[0010] Compliance with cGMP and product approval are some regulatory challenges. Extensive documentation, validation, and quality control measures may be required to comply with regulatory standards. Navigating complex regulatory pathways and ensuring each production batch meets regulatory standards represent some current challenges.

[0011] High initial investment and operational costs are some of the economic setbacks. Significant capital expenditure is required for building and equipping cGMP-compliant facilities. High operational costs, including raw materials, labor, and quality control, may be challenging, especially during the initial scale-up phase.

[0012] Current processes and strategies for scaling cell production may include bioreactor systems, modular and single-use systems, process automation, and / or quality control measures. However, these methods still face significant challenges in achieving efficient, consistent, and scalable cell production.

[0013] Examples of bioreactor systems include stirred-tank bioreactors, wave bioreactors, hollow-fiber bioreactors, and single-use bioreactors. Stirred tank bioreactors may be used for suspension and / or adherent cell cultures, and may be equipped with control systems for monitoring. These systems are may be and suitable for high-density cultures. However, mechanical stress and potential for non-uniform mixing are drawbacks to this type of bioreactor. Wave bioreactors utilize rocking motion to provide mixing and aeration, and may be suitable for suspension cultures. They may reduce shear stress and be scalable. However, scalability by unit increase and limitations to certain cell types are disadvantages to this bioreactor system. Hollow-fiber bioreactors may provide surface area for adherent cell growth within a compact system. These systems may support nutrient and oxygen exchange and high- density cultures, but experience complexity in scaling and potential shear stress. Single-use bioreactors are disposable systems that reduce contamination risk and cleaning and sterilization needs. Flexibility, reduced downtime, and cost-effectiveness for small to medium-scale production are advantages to the single-use bioreactor system. However, waste generation and limitation to specific scales are disadvantages to this bioreactor system.

[0014] 2D cell culture methods involve growing cells in a monolayer on flat, rigid surfaces such as plastic dishes, plates, or flasks. These methods are used due to their simplicity, lower cost, and relative ease of use. However, 2D cultures fail to replicate the 3D environment of tissues and organs, leading to significant differences in cell morphology, behavior, and function. Cells in 2D cultures often exhibit altered gene expression and reduced functionality compared to their in vivo counterparts, resulting in limited predictive power for drug efficacy and toxicity studies.

[0015] Developed 3D cell culture systems, and / or spheroids, may address some limitations of 2D cultures. Cells may grow in three dimensions, better mimicking the in vivo environment. Where 3D cultures may improve cell-cell or cell-matrix interactions, they often face challenges related to nutrient and oxygen diffusion, especially in larger spheroids, leading to necrotic cores. Additionally, the formation and consistency of 3D spheroids may be variable, complicating experimental reproducibility.

[0016] Organoids are 3D structures derived from stem cells or primary tissues that selforganize into miniaturized versions of organs, retaining certain features of their in vivo counterparts. Organoids have become tools for studying organ development, disease mechanisms, or drug responses. However, their complexity and high cost limit scalability and widespread adoption. Additionally, organoids often lack vascularization, which can be important for maintaining long-term cell viability and / or function.

[0017] Microfluidic systems and organ-on-a-chip devices may use microfabrication techniques to create channels and chambers that simulate the microenvironment and fluid flow conditions of tissues and organs. These systems may provide dynamic culturing conditions and support the integration of multiple cell types, offering more physiologically relevant models. However, they require specialized knowledge and equipment, making them technically complex and expensive. Scalability remains a significant challenge for high-throughput applications.

[0018] 3D bioprinting can involve using a printer to create tissue structures by depositing layers of bioink containing cells or biomaterials. Bioprinting may offer relative precision and customization, allowing for creation of tissue architectures. Despite this, bioprinting is expensive, technically demanding, and often limited by the availability of suitable bioinks and the complexity of replicating functional tissues. Competitive technologies for biologies production may include CHO (Chinese Hamster Ovary) cells, HEK293 (Human Embryonic Kidney) cells, and Vero cells (African Green Monkey Kidney Cells). CHO cells may produce therapeutic proteins and monoclonal antibodies. However, ensuring consistent glycosylation patterns and maintaining cell viability during scale-up remain difficult. HEK293 advantages may include gene therapy vectors and production of therapeutic proteins, but difficulties achieving high virus titers and ensuring genetic stability are setbacks. Vero cells may help produce viral vaccines, but issues preventing contamination and optimizing media formulations are disadvantages.

[0019] The limitations of current cell culture methods and the challenges associated with scaling cell and biologic production underscore the need for improved technologies that may provide more physiologically relevant models, enhance scalability, and reduce costs.

[0020] Citation or identification of any document in this application is not an admission that such a document is available as prior art to the present disclosure.

[0021] SUMMARY

[0022] One or more embodiments of the disclosure may address one or more of the aforementioned problems. Certain embodiments according to the disclosure may provide a modular cell culture system, which may comprise at least one interconnecting porous hydrogel block (IPHB); wherein the IPHB(s) may comprise a biocompatible hydrogel matrix with an interconnected network of cavities (e.g. microchannel, pore, and / or chamber); wherein the IPHB(s) may be configured to interconnect horizontally and / or vertically, e.g. to form a customizable and / or expandable culture environment; and one or more microchannels may be configured to provide pathway(s) for nutrient delivery, gas exchange, and / or waste removal. Some embodiments may be configured to mimic the extracellular matrix (ECM) of native tissues, e.g. supporting natural cell adhesion, growth, and / or differentiation. Some embodiments may be configured to be oxygen-permeable and / or swellable, which could be based on salinity, e.g. of perfused liquid(s). Still yet, the microchannels within at least one IPHB may be configured to form a pseudo-vasculature network, e.g. facilitating the efficient transport of nutrients and / or removal of waste products. Yet again, at least one IPHB may be to support both static and perfusion-based cultures, e.g. allowing for controlled delivery of nutrients, gases, and / or mechanical stimuli. Still yet further, at least one IPHB may be configured to enable the co-culture of multiple cell types within the same system, e.g. to facilitate the formation of complex tissue structures and / or organ components. At least one IPHB may be configured to be enzymatically dissolved in certain embodiments, e.g. to release cells, such as for downstream applications, which may preserve cell viability and / or functionality. In some embodiments, the continuous polymeric matrix material is enzymatically dissolvable. Some embodiments may be configured for histological and / or immunohistochemical analysis, e.g. to allow for detailed examination of tissue architecture and / or cellular interactions. Some embodiments may be configured to reduce media and / or plasticware consumption compared to traditional cell culture methods, e.g. lowering overall production costs.

[0023] In another aspect, the present disclosure provides a system for personalized medicine, which may comprise at least one IPHB configured to support the co-culture of patient-specific cells; and / or the capability to form patient-specific tissue model(s) for drug testing and / or therapy optimization. Some embodiments may be configured to allow for the comparison of healthy and / or diseased tissues, e.g. to identify specific drug targets and / or optimize treatment protocols for individual patients.

[0024] In another aspect, the present disclosure provides a method for producing organized tissue structure(s) and / or functional organ components), which may comprise seeding cells into the microchannels of the at least one IPHB; interconnecting at least one IPHB horizontally and / or vertically to expand the culture environment; and providing nutrients, gases, and / or mechanical stimuli, such as through static or perfusion-based cultures, e.g. to promote cell growth and / or tissue organization. Some embodiments may be configured to support the formation of a pseudo-vasculature network within the tissue constructs. Some embodiments may be configured to be oxygen-permeable and / or swellable, e.g. enhancing cell viability and / or function. Some embodiments may comprise enzymatically dissolving hydrogel matrix, e.g. to release cells, such as for downstream application(s). In some embodiments, co-culture of multiple cell types within the IPHB(s) may be configured to facilitate development of complex tissue structure(s) and / or organ components). Some embodiments may be configured to enable the production of high-density cell cultures and / or high yields of biologies, e.g. improving cost-effectiveness and / or scalability. The modular and / or expandable nature of the IPHB(s) and / or embodiments herein may be configured to support small-scale research and / or large-scale industrial applications).

[0025] In another aspect, the present disclosure provides a method for enhancing drug discovery and / or screening, which may comprise using at least one IPHB to create at least one high-throughput and / or physiologically relevant model for preclinical study(ies); and assessing drug efficacy and / or toxicity in the tissue model(s), such as those formed within the IPHB(s). Further, the IPHB(s) may be configured to enable analysis of cellular response(s) and / or interaction(s), e.g. providing insight(s) into experimental outcome(s).

[0026] In certain embodiments, a modular cell culture system may comprise a plurality of IPHBs. One or more blocks may comprise a three-dimensional (3D) continuous polymeric matrix with a network of microporous cavities. One or more blocks may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal.

[0027] Some embodiments may be configured to mimic at least one ECM.

[0028] Some embodiments may be configured to match at least one physical property of an ECM.

[0029] In some embodiments, one or more physical properties may comprise porosity, stiffness, hardness, capacitance, elasticity, elastic modulus, density, conductivity, thermal conductivity, electric conductivity, electric charge, solubility, hydrophilicity, and / or lipophilicity.

[0030] In some embodiments, at least one block may be oxygen permeable.

[0031] In some embodiments, at least one block may be oxygen permeable based on the salinity of at least one perfusate.

[0032] In some embodiments, at least one block may be swellable.

[0033] In some embodiments, at least one block may be swellable based on the salinity of at least one perfusate.

[0034] In some embodiments, one or more pathways may be configured to mimic pseudovasculature.

[0035] Some embodiments may be configured to permit exposure of one or more cavities to at least one nutrient, gas, mechanical stimulus, static solution, and / or perfusion.

[0036] In some embodiments, at least one cavity may comprise one or more channels, chambers, and / or pores.

[0037] In some embodiments, at least one block may be configured to be enzymatically dissolvable.

[0038] Some embodiments may be configured for histological and / or immunohistochemical study.

[0039] Some embodiments may be configured to cultivate one or more cell types when seeded with at least one cell of each type. Some embodiments may be configured to permit co-cultivation of a plurality of cell types within at least one block.

[0040] In some embodiments, one or more cells may be derived from a specimen, human, subject, tissue, organ, and / or biopsy. The biopsy could be from the subject or otherwise. The subject could be a human or a nonhuman. The subject could be a patient. Multiple specimens could be from the same and / or different sources; examples include multiple specimens from the same biopsy, different biopsies from the same subject (e.g. including differing in the location and / or time biopsied), and / or different subjects. Additional permutations will be readily apparent to those of ordinary skill in the art.

[0041] In some embodiments, at least one cavity may be configured for communication between two or more interconnected blocks in one or more vertical, horizontal, and / or orthogonal axes relative to gravity.

[0042] In certain embodiments, a modular cell culture method may comprise seeding at least one IPHB with at least one cell type. One or more blocks may comprise a 3D continuous polymeric matrix with a network of microporous cavities. One or more blocks may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal.

[0043] In some embodiments, at least one cell may be isolated from at least one subject. One or more subjects may comprise a human, mammal, animal, bird, reptile, amphibian, and / or fish. At least one cell may be isolated from one or more organs and / or tissues.

[0044] Some embodiments may further comprise cultivating the cells into a personalized model and subjecting the model to analysis to guide diagnosis and / or therapy tailored for the subject.

[0045] In certain embodiments, a modular cell culture IPHB may comprise a 3D continuous polymeric matrix with a network of microporous cavities. The block may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal.

[0046] These and other aspects, objects, features, and advantages of the example embodiments will become apparent to those having ordinary skill in the art upon consideration of the following detailed description of example embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] An understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure may be utilized, and the accompanying drawings of which:

[0048] FIG. 1 illustrates two separate interconnecting porous hydrogel blocks (IPHBs) in accordance with certain embodiments;

[0049] FIG. 2 illustrates three interconnected IPHBs in accordance with certain embodiments; and

[0050] FIG. 3 illustrates a modular cell culture method in accordance with certain embodiments.

[0051] The figures herein are for illustrative purposes only and are not necessarily drawn to scale.

[0052] DETAILED DESCRIPTION OF THE EXAMPLE EMBODIMENTS

[0053] The disclosure now will be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments are shown. Indeed, what is claimed may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. As used in the specification, and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise.

[0054] Certain presently disclosed embodiments relate to interconnecting porous hydrogel blocks (IPHBs). IPHBs and embodiments herein represent novel and advanced platforms designed to transform the production of mammalian cells and biologies. This disclosure addresses the limitations of traditional cell culture methods and current bioreactor technologies, e.g. by providing scalable, physiologically relevant, and / or cost-effective solutions for cell and / or biologic production. IPHBs and embodiments herein may enable the formation of organized tissue structure(s) and / or functional organ components), e.g. offering significant advancements in drug discovery, biotherapeutic development, regenerative medicine, and / or personalized therapies.

[0055] Figure 1, for instance, illustrates two (2) separate IPHBs 1 in accordance with certain embodiments. Each of these IPHBs include a top surface 12, a bottom surface 14, and at least one side edge 16. The particular IPHBs 1 shown in Figure 1 include at least one interlockingmale component 50 and at least one interlocking-female component 60. Figure 2 illustrates three (3) interconnected IPHBs 1 in accordance with certain embodiments. The IPHBs 1 shown in Figure 2 each include a first interlocking-male component 51, a second interlocking-male component 52, a first interlocking-female component 61, and a second interlocking-female component 62. IPHBs 1 may interconnect via first interlocking-male component 51 or a second interlocking-male component 52 interfacing with a first interlocking-female component 61 or a second interlocking-female component 62, or vice versa. In certain embodiments, one or more interlocking-male components (50, 51, 52) comprise a protrusion. In certain embodiments, one or more interlocking-female components (60, 61, 62) comprise a recess. In certain embodiments, one or more interlocking-female components (60, 61, 62) is a recess configured complimentary to a protrusion of one or more interlocking-male components (50, 51, 52). Persons of ordinary skill will readily appreciate additional variations. For example, one or more interlocking-male components (50, 51, 52) may protrude from an IPHB l’s top surface 12, bottom surface 14, one or more side edges 16, or any combination thereof. One or more interlocking-female components (60, 61, 62) may recess into an IPHB 1 ’s top surface 12, bottom surface 14, one or more side edges 16, or any combination thereof. One or more interlocking-male components (50, 51, 52) may comprise a quadrangular protrusion, or any other angled or rounded shape, or any combination thereof. One or more interlocking-female components (60, 61, 62) may comprise a quadrangular recess, or any other angled or rounded shape, or any combination thereof.

[0056] Despite advancements in cell culture technologies, several gaps and barriers hinder the successful translation of research into therapies. Most current in vitro models lack functional vasculature, limiting nutrient and oxygen supply in larger constructs, which is important for maintaining long-term cell viability and function. Existing models often fail to replicate extracellular matrix (ECM) composition, mechanical properties, and cellular interactions of native tissues, leading to limited predictive power for drug efficacy and toxicity studies. Many in vitro models are challenging to scale up for high-throughput screening or industrial applications. High costs and technical complexity limit the accessibility and widespread adoption of technologies like organ-on-a-chip devices and bioprinting. Variability in the formation and behavior of three-dimensional (3D) cultures and organoids may lead to inconsistent experimental results. IPHBs and embodiments herein offer solutions, e.g. by providing a modular, scalable, and / or tissue-mimetic platform to address these limitations, enabling more accurate and / or reliable preclinical models, accelerating therapeutic development, and / or reducing the need for animal testing.

[0057] Embodiments herein feature unique modular design, e.g. allowing for horizontal and / or vertical interconnection. The ability to connect multiple IPHBs may provide researchers with flexibility to scale up experiments and / or production processes, e.g. without significant infrastructural changes and / or increased costs. Embodiments herein, including modular systems and / or IPHBs that may be interconnected horizontally and / or vertically or otherwise, e.g. to expand and / or customize cell culture environment, represent novel features that go beyond traditional static cultures and existing bioreactor systems. Where modularity might exist in bioreactor design, the unique features and / or implementations herein may allow more seamless expansion and / or configuration, e.g. to suit various experimental needs. Traditional bioreactors, such as stir-tank or wave bioreactors, do not offer this level of flexibility and scalability, which are important for adapting to different cell culture requirements and scaling production efficiently.

[0058] Among innovative aspects of IPHBs and embodiments herein are implementations comprising integrated pseudo-vasculature network(s) of interconnected cavities (e.g., microchannels within the hydrogel matrix). Such network(s) may facilitate efficient nutrient delivery, gas exchange, and / or waste removal, which can be important for maintaining cell viability and / or function, especially in large, dense tissue constructs. The microchannels in some embodiments may be aligned and / or expanded by connecting multiple IPHBs horizontally and / or vertically, e.g. allowing for customizable configurations and / or scalability. Unlike traditional 2D cultures or simple 3D spheroids, IPHBs and embodiments herein may help cells receive a more consistent supply of essential nutrients and / or oxygen, e.g. more closely mimicking in vivo conditions. The incorporation of an interconnected microchannel network within hydrogel matrix to form pseudo-vasculature system is among innovative features herein and may offer a solution to the common problem of nutrient and oxygen delivery in dense cell cultures.

[0059] Certain embodiments and / or IPHBs herein may support static and / or perfusion-based cultures, e.g. allowing researchers to simulate dynamic in vivo conditions. In some embodiments the ability to perfuse media may be under controlled pressures and / or flow rates, among features representing a significant improvement over traditional static cultures (which often suffer from limited nutrient diffusion and / or waste accumulation). The dynamic environment of certain embodiments herein may promote more natural cell behavior, organization, and / or functionality. Where microfluidic systems and / or organ-on-a-chip devices may provide dynamic conditions, they are typically more complex and less scalable.

[0060] The hydrogel matrix used in the IPHB(s) of certain embodiments herein may be designed to closely mimic the ECM found in native tissues. This biocompatible material of certain embodiments herein may support natural cell adhesion, growth, and / or differentiation, e.g. providing a more physiologically relevant environment compared to the rigid surfaces of traditional plastic culture dishes. Additionally, certain embodiments comprising oxygen permeability and / or swellable properties, which could be based on salinity (e.g. of perfused liquid(s)) may enhance cell viability and / or function.

[0061] Certain embodiments and IPHBs may enable the co-culture of multiple cell types within the same implementation and / or unit, e.g. facilitating the formation of complex tissue structure(s) and / or organ components). Features herein are particularly valuable for creating more accurate disease models and / or studying cell-cell interaction(s), e.g. in more controlled environments). Traditional cell culture methods often struggle with maintaining multiple cell types in a single culture due to differences in growth requirements) and / or condition(s).

[0062] The design of certain embodiments and / or IPHBs herein may allow for extensive analytical capabilities that are not feasible with many existing systems. Researchers may enzymatically dissolve hydrogel matrix, for example, to release cells such as for profiling; and / or may fix and / or section the IPHB(s) for histological and / or immunohistochemical analysis. In some implementations, the IPHB(s) may be non-degradable. Variability and / or flexibility of features herein may enable more detailed examination of tissue architecture, cellular interaction(s), and / or biochemical process(es), such as by providing more comprehensive insight(s), e.g. into experimental outcome(s). Traditional methods often require separate, complex procedures for analyses, whereas embodiments herein may integrate capabilities. Electrophysiological assessments) and / or biochemical analysis of metabolites and / or signaling molecules may also be used with certain IPHBs and / or embodiments herein, such as to analyze cultured tissue(s). This may provide valuable insight(s), e.g. into experimental outcome(s), and / or support the development of new therapeutic(s).

[0063] In some embodiments, gentle harvesting facilitated by enzymatic dissolution of hydrogel matrix may preserve cell viability and / or functionality, which can be important for sensitive cell types like stem cells. Features herein offer significant advantage(s) over traditional methods that often involve mechanical disruption, which may damage sensitive cell types, such as stem cells. Embodiments herein promote high-quality, functional cells, which may be obtained e.g. for downstream applications), such as therapeutic use.

[0064] IPHBs and embodiments herein may significantly reduce the consumption of media and / or plasticware compared to traditional cell culture methods. The high-density cultures supported by implementations herein may lead to higher yields of cells and / or biologies per unit volume, e.g. improving cost-effectiveness and / or making advanced cell culture technologies more accessible. The closed system design of certain embodiments may also minimize the risk of contamination, e.g. reducing the need for extensive sterility measures and / or associated costs.

[0065] The field of the disclosure may include development, production, and / or application of certain embodiments and / or IPHBs for various purposes e.g. in the life sciences, which can emphasize their potential to transform current practices and / or drive innovation such as in cell and / or biologic production. IPHBs and embodiments herein may be suitable for a wide range of applications, including drug discovery and / or screening, biotherapeutic development, regenerative medicine, and / or personalized medicine.

[0066] IPHBs and embodiments herein may provide high-throughput and / or physiologically relevant platform(s) for drug discovery and / or screening. The ability to create complex tissue model(s) and / or simulate dynamic in vivo condition(s) may enhance the accuracy and / or reliability of preclinical study(ies), e.g. improving the prediction of drug efficacy and / or toxicity. Such advancement may reduce the need for animal testing and / or accelerate the development of new therapeutic(s).

[0067] IPHBs and embodiments herein may support the production of high-quality biologic(s), such as one or more therapeutic proteins, antibodies, and / or exosomes. Their scalable and cost- effective features may make them suitable for industrial application^), e.g. enhancing the efficiency and / or yield of biotherapeutic production.

[0068] The ability to form organized tissue structure(s) and / r functional organ components) may make embodiments herein valuable for regenerative medicine. They may be used to generate tissue graft(s), study tissue regeneration process(es), and / or develop regenerative therapy(ies).

[0069] Embodiments herein may enable the creation of patient-specific tissue model(s), e.g. for personalized drug testing and / or therapy optimization. By comparing healthy and / or diseased tissue(s), research(ers) may identify specific drug target(s) and / or optimize treatment protocol(s), such as for individual patient(s), e.g. advancing the field of personalized medicine.

[0070] IPHBs and embodiments herein represent transformative approach to cell and / or biologic production, e.g. by addressing limitations of existing technologies. The novel features herein can enhance the physiological relevance, scalability, and / or efficiency of in vitro models, and may enable more accurate and / or reliable preclinical studies. They may accelerate the development of new therapeutic(s), reduce the need for animal testing, and / or advance personalized medicine, such as by providing patient-specific tissue model(s), e.g. for drug testing and / or therapy optimization.

[0071] Certain embodiments may be configured to mimic and / or resemble in vivo conditions associated with a natural cell and / or tissue of interest. For example, a continuous polymeric matrix material may mimic a natural tissue of interest by matching one or more physical properties of said tissue within about 20% of a measurable goal or threshold. Examples may include deviation by about 15%, 10%, 8%, 5%, 3%, or 1%, from the natural tissue of interest. The one or more physical properties may include, for example, softness and / or tension strength, and / or elasticity. One or more physical properties may comprise porosity, stiffness, and / or elastic modulus. Additionally or alternatively, the network of microporous cavities (e.g. channels and / or chambers) may be structured to mimic the morphology of a natural tissue of interest, such as by varying the geometry and / or dimensions of a network of microporous channels and / or chambers to mirror the morphology of the natural tissue of interest. In certain examples, a 3D network of microporous channels and / or chambers defines a 3D scaffolding for propagation of one or more cell types of interest. In accordance with certain embodiments, an average diameter of one or more microporous cavities may comprise from about 100 to about 800 microns. The minimum diameter of one or more microporous cavities may comprise 100, 120, 150, 180, 200, 220, and / or 250 microns, in certain embodiments. The maximum diameter of one or more microporous cavities may comprise 800, 780, 750, 720, 700, 680, 650, 620, 600, 580, 550, 520, 500, 480, 450, 420, 400, 380, 350, 320, 300, 280, and / or 250 microns, in certain embodiments. Additionally or alternatively, microporous channels and / or chambers may comprise at least about 40% by volume of the 3D macrostructure of an IPHB. In certain embodiments, microporous cavities may comprise at least about 40, 50, 60, and / or 70% by volume of the 3D macrostructure. In certain embodiments, microporous cavities may comprise at most about 90, 85, 80, 75, and / or 70% by volume of the 3D macrostructure. Embodiments herein may be suitable for a variety of applications, such as producing and / or growing cell cultures, bacteria cultures, yeast cultures, biologies, exosomes, extracellular vesicles, growth factors, monoclonal antibodies, peptides, proteins, viral particles, oligonucleotides, and / or organelles; organoid formation, plant growth, drug delivery, tissue formation, ex vivo modeling, electrical conduction, wound healing, cellular reprogramming, filtration, optics, and / or microfluidics; helping construct and / or form a custom network of microchannels, custom scaffold architecture, custom ECM derived scaffold, dissolvable hydrogel, custom tissue, custom tissue formation, and / or custom configuration; accepting patient cells; and / or microenvironment manipulation.

[0072] In accordance with certain embodiments of the disclosure, the continuous polymeric matrix material may be generally non-degradable. In certain examples, the cells and / or tissue produced in the IPHB may need to be flushed out of the interior network of the network of microporous channels and / or chambers for further analysis, purification, and / or development. Additionally or alternatively, the continuous polymeric matrix material may be selectably degradable (e.g., enzymatically dissolved). For example, hydrogel formulations may be rendered biodegradable, such as by insertion of enzyme-sensitive sequences and / or utilization of native matrix-derived compounds. For example, the continuous polymeric matrix material may comprise a selectably degradable hydrogel material comprising one or more degradable polymers, such as one or more biopolymers derived from a living organism. The one or more biopolymers derived from a living organism, for example, may comprise a polynucleotide, polysaccharide, polypeptide, or any combination thereof. In accordance with certain embodiments, the one or more biopolymers may comprise collagen, gelatin (e.g., porcine, bovine [cow], ovine [sheep], shellfish, fish, etc.), laminin, alginate, glycosaminoglycans, oligonucleotides (e.g., DNA, RNA), carbohydrates, lipids, cellulose, alginate, and / or proteins that can be gently degraded, such as with the use of specific enzymes, ionic solvents, neutral detergents, weak acids, and / or peroxides to disrupt the biopolymer chains. In accordance with certain embodiments, the one or more biopolymers may comprise degradable monomers comprising esters, such as hydroxybutyrate, lactic acid, glycolic acid, and / or caprolactone; anhydrides, such as adipic acid and / or sebacic acid; saccharides, such as cellulose, alginate, pectin, dextrin, chitosan, hyaluronan, chondroitin sulfate, and / or heparin; proteins; nucleotides, such as DNA and / or RNA; peptides, such as collagen, gelatin, silk, and / or fibrin; urethanes; phosphates; carbonates; and / or vinyl chlorides. Additionally or alternatively, the biopolymer may comprise poly(lactic-co-glycolic acid) (PLGA), polyethylene glycol (PEG), and / or other suitable hydrogels. In accordance with certain embodiments, the selectably degradable hydrogel material may comprise a synthetic polymer, such as a polyester, a polyanhydride, a polycarbonate, a polyurethane, a polyphosphate, or any combinations) thereof. The continuous polymeric matrix material, in accordance with certain embodiments, may comprise a 3D crosslinked polymer network, a non-crosslinked polymer network, or a combination thereof.

[0073] In certain embodiments, one or more biopolymers may comprise anywhere between approximately 10 and 100% of the dry weight of an IPHB and / or its continuous polymeric matrix material. In certain embodiments, the minimum dry weight of an IPHB and / or its continuous polymeric matrix material to consist of one or more biopolymers may be about 10, 15, 20, 25, 30, 35, 40, 45, and / or 50%. In certain embodiments, the maximum dry weight of an IPHB and / or its continuous polymeric matrix material to consist of one or more biopolymers may be about 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, and / or 50%.

[0074] In certain embodiments, a continuous polymeric matrix material may comprise a swellable hydrogel material. The swellable hydrogel material may comprise a radically mediated reaction product of at least a first monomer including acrylate or methacrylate functional group(s) and a second monomer or oligomer including at least two (2) free-radically polymerizable functional groups. For example, the at least two (2) free-radically polymerizable functional groups may independently from each other comprise an acrylate or methacrylate group, an allylic group, an alkynyl, a vinyl nitrile, a vinyl ether, a vinyl ester, a vinyl amide, a styrenic group, a maleate group, a fumarate group, and / or a norbomene group. In accordance with certain embodiments, at least one of the first monomer and / or the second monomer may comprise polyethylene glycol functionality (e.g., — O(C2H4O)nH; where n has a value from 1 to 100), polypropylene glycol functionality (e.g., — O(C3HeO)nH; where n has a value from 1 to 100), and / or glycerol functionality incorporated into a backbone of the monomer and / or grafted onto the monomer as a side-chain or a component of a side chain. By way of example, at least one of the first monomer and / or second monomer may comprise 2-Hydroxyethyl acrylate (HEA), Poly(ethylene glycol) methyl ether acrylate (MPEGA), N-Methyl acetamide (NMA), or Poly(ethylene glycol) diacrylate (PEGDA). In accordance with certain embodiments, non-limiting examples of non-degradable monomers that may be utilized in hydrogel materials may include polyolefins (e.g., ethylene, propylene), styrene, nylon (e.g., amides), and / or acrylics. In accordance with certain embodiments, non-limiting examples of degradable monomers that may be utilized in hydrogel materials may include esters (e.g., hydroxybutyrate, lactic acid, glycolic acid, caprolactone), anhydrides ( e.g., adipic acid, sebacic acid), saccharides (e.g., cellulose, alginate, pectin, dextrin, chitosan, hyaluronan, chondroitin sulfate, heparin), proteins, nucleotides (e.g., DNA, RNA), peptides (e.g., collagen, gelatin, silk, fibrin), urethanes, phosphates, carbonates, and / or vinyl chlorides. Additionally or alternatively, a third monomer comprising a cross-linking agent may be incorporated in continuous polymeric matrix material. Additionally or alternatively, the swellable hydrogel material may comprise one or more natural polymers, such as plant-derived polymers (e.g., cellulosic- polymers) and / or animal-derived polymers. A natural polymer herein may comprise laminin. Further, a gelatin herein may comprise porcine, bovine (cow), ovine (sheep), shellfish, and / or fish gelatin.

[0075] In certain embodiments, one or more thermoplastic polymers may comprise between about 10 to 100% of the dry weight of an IPHB and / or its continuous polymeric matrix material. In certain embodiments, the minimum dry weight of an IPHB and / or its continuous polymeric matrix material to consist of one or more thermoplastic polymers may be about 10, 15, 20, 25, 30, 35, 40, 45, and / or 50%. In certain embodiments, the maximum dry weight of an IPHB and / or its continuous polymeric matrix material to consist of one or more thermoplastic polymers maybe about 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, and / or 50%.

[0076] In accordance with certain embodiments, an interface between the network of microporous cavities (e.g., channels and / or chambers) and the continuous polymeric matrix material may comprise a coating of at least one compatibilizer selected to promote adhesion of at least one cell of interest. This coating may be applied subsequent to IPHB formation. By way of example, the coating comprising the compatibilizer(s) may comprise a biological coating, including, for example, collagen I (e.g., from or for human mesenchymal stem cells {MSCs} [e.g., from or for adipose, bone barrow, and / or umbilical cord tissues], human neonatal dermal fibroblasts, human adult dermal fibroblasts, human keratinocytes, human myocytes, human osteoblasts, human osteocytes, human chondrocytes, bovine myocytes, porcine hepatocytes, porcine chondrocytes, porcine osteocytes, and / or equine muscle derived stem cells); laminin I (e.g., from or for human induced pluripotent stem cells {IPSCs} and / or mouse dorsal root ganglia); hyaluronan (e.g., from porcine hepatocytes and / or human dermal adult fibroblasts); gelatin (e.g., from human MSCs [e.g., from or for adipose, bone marrow, and / or umbilical cord tissues], human neonatal dermal fibroblasts, human adult dermal fibroblasts, human keratinocytes, human myocytes, human osteoblasts, human osteocytes, human chondrocytes, human CD8+ T cells, Human CD4+ T cells, human macrophages, bovine myocytes, porcine hepatocytes, porcine chondrocytes, porcine osteocytes, and / or equine muscle derived stem cells); fibrin (e.g., from or for human keratinocytes); fibronectin (e.g., from or for human MSCs [e.g., from or for adipose, bone marrow, and / or umbilical cord tissues], human neonatal dermal fibroblasts, human adult dermal fibroblasts, human keratinocytes, human osteoblasts, human osteocytes, and / or human chondrocytes); or any combination(s) thereof.

[0077] By way of example, in certain embodiments one or more IPHBs may be seeded and / or configured to enable cell growth of one or more of the following: human stem cells, such as human Wharton’s Jelly cells, human umbilical cord derived MSCs, human bone marrow derived MSCs, human adipose derived MSCs, human skin derived IPSCs, human blood cell derived IPSCs, human CD4+ T cells, and human CD8+ T cells; primary mammalian cells, such as HepG2 cells or other liver carcinoma cells, human adult dermal fibroblasts, human neonatal dermal fibroblasts, human adult keratinocytes, mouse dorsal root ganglia or other primary neural cells, bovine myocytes, porcine hepatocytes, porcine chondrocytes, porcine osteocytes, equine muscle derived MSCs, snail cells, and human macrophages; and immortalized mammalian cell lines, such as UB-OC2 or mouse cochlear epithelium cells, human myoblastoma or other muscle tumor cells, PC3 or other prostate cancer cells, CHO or Chinese hamster ovary cells, HEK293 or other human embryonic kidney cells, SHSY5Y or other neuronal tumor cells, PANC-1 or other human pancreatic cancer cells, HeLa or other cervical cancer cells, A549 or other lung cancer cells, and A673 or other muscle cancer cells; and primary plant cells, such as rosemary, tobacco, and tomato.

[0078] In certain embodiments, a method of generating mature mammalian tissue may comprise providing an initial scaffolding comprising at least one IPHB that comprises a 3D macrostructure defined by a continuous polymeric matrix material with a network of microporous cavities (e.g. pores, channels and / or chambers), seeding the IPHB(s) with one or more types of adherent cells, and allowing the cells to propagate and form mature mammalian tissue within the IPHB(s). Some embodiments may comprise a first IPHB and a second IPHB, interconnecting at least the first IPHB and the second IPHB to expand an initial scaffolding to expand the culture environment and generate larger mature mammalian tissue. Adherent cells may comprise primary mammalian cell lines, MSCs, IPSCs, immortalized cell lines, or combinations thereof. The IPHB(s) may be seeded with at least two different cell types, and co-cultured to allow cells to interact and regulate into mature tissue(s). The IPHB(s) may be perfused with culture media, e.g. to provide consistent nutrient delivery, and / or waste removal, which could be throughout the generation process. The IPHB(s) in embodiments herein may be configured to be enzymatically dissolved, e.g. to retrieve tissue. Some embodiments may be configured to mimic the native physiological conditions, such as from a biological, chemical, electrical, genetic, physical, structural, and / or mechanical perspective, e.g. facilitating the generation of physiologically relevant tissue(s).

[0079] Tissue generated from any embodiment herein may be harvested, and for example may be suitable for application(s) in research modeling, drug testing, and / or the development of tissue graft(s), such as for treating patient(s).

[0080] In certain embodiments, a method for producing organized tissue structure(s) and / or functional organ components) may comprise seeding cells into the microchannels of at least one IPHB; interconnecting a plurality of IPHBs horizontally and / or vertically to expand the culture environment; and providing nutrients, gases, and / or mechanical stimuli, such as through static and / or perfusion-based culture(s), e.g. to promote cell growth and / or tissue organization. Some embodiments may be configured to support the formation of a pseudo-vasculature network, e.g. within tissue constructs). Some embodiments may be configured to be oxygen- permeable and / or swellable, e.g. enhancing cell viability and / or function. Some embodiments may be configured to be enzymatically dissolved, e.g. to release cells, such as for downstream applications). Multiple cell types within the IPHB(s) may be co-cultured, e.g. to facilitate the development of complex tissue structure(s) and / or organ components). Some embodiments may enable the production of high-density cell culture(s) and / or high yields of biologic(s), e.g. improving cost-effectiveness and / or scalability. The modular and / or expandable nature of certain embodiments may support small-scale research and / or large-scale industrial application(s).

[0081] In certain embodiments, a method for enhancing drug discovery and / or screening may comprise using at least one IPHB to create one or more high-throughput and / or physiologically relevant model(s), e.g. for preclinical study(ies); and / or assessing drug efficacy and / or toxicity in tissue model(s), such as those formed within the IPHB(s). Some embodiments may enable more detailed analysis of cellular response(s) and / or interaction(s), e.g. providing more comprehensive insight(s) into experimental outcome(s).

[0082] In certain embodiments, a modular cell culture system may comprise a plurality of IPHBs. One or more blocks may comprise a 3D continuous polymeric matrix with a network of microporous cavities. One or more blocks may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal. Some embodiments may be configured to mimic at least one ECM.

[0083] Some embodiments may be configured to match at least one physical property of an ECM.

[0084] In some embodiments, one or more physical properties may comprise porosity, stiffness, hardness, capacitance, elasticity, elastic modulus, density, conductivity, thermal conductivity, electric conductivity, electric charge, solubility, hydrophilicity, and / or lipophilicity.

[0085] In some embodiments, at least one block may be oxygen permeable.

[0086] In some embodiments, at least one block may be oxygen permeable based on the salinity of at least one perfusate.

[0087] In some embodiments, at least one block may be swellable.

[0088] In some embodiments, at least one block may be swellable based on the salinity of at least one perfusate.

[0089] In some embodiments, one or more pathways may be configured to mimic pseudovasculature.

[0090] Some embodiments may be configured to permit exposure of one or more cavities to at least one nutrient, gas, mechanical stimulus, static solution, and / or perfusion.

[0091] In some embodiments, at least one cavity may comprise one or more channels, chambers, and / or pores.

[0092] In some embodiments, at least one block may be configured to be enzymatically dissolvable.

[0093] Some embodiments may be configured for histological and / or immunohistochemical study.

[0094] Some embodiments may be configured to cultivate one or more cell types when seeded with at least one cell of each type.

[0095] Some embodiments may be configured to permit co-cultivation of a plurality of cell types within at least one block.

[0096] In some embodiments, one or more cells may be derived from a specimen, human, subject, tissue, organ, and / or biopsy. The biopsy could be from the subject or otherwise. The subject could be a human or a nonhuman. The subject could be a patient. Multiple specimens could be from the same and / or different sources; examples include multiple specimens from the same biopsy, different biopsies from the same subject (e.g. including differing in the location and / or time biopsied), and / or different subjects. Additional permutations will be readily apparent to those of ordinary skill in the art.

[0097] In some embodiments, at least one cavity may be configured for communication between two or more interconnected blocks in one or more vertical, horizontal, and / or orthogonal axes relative to gravity.

[0098] In certain embodiments, a modular cell culture method may comprise seeding at least one IPHB with at least one cell type. One or more blocks may comprise a 3D continuous polymeric matrix with a network of microporous cavities. One or more blocks may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal.

[0099] In some embodiments, at least one cell may be isolated from at least one subject. One or more subjects may comprise a human, mammal, animal, bird, reptile, amphibian, and / or fish. At least one cell may be isolated from one or more organs and / or tissues.

[0100] Some embodiments may further comprise cultivating the cells into a personalized model and subjecting the model to analysis to guide diagnosis and / or therapy tailored for the subject.

[0101] In certain embodiments, a modular cell culture IPHB may comprise a 3D continuous polymeric matrix with a network of microporous cavities. The block may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal.

[0102] Described herein are modular cell culture systems, methods, and IPHBs. A block may comprise a 3D continuous polymeric matrix with a network of microporous cavities, and may be configured to interconnect with at least one other block. One or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal. A system may comprise at least one IPHB so configured. A method may comprise seeding at least one IPHB so configured with at least one cell type.

[0103] Turning to Figure 3, illustrated is an example embodiment of a modular cell culture method 300. In certain embodiments, method 300 may comprise step 310 of seeding and optionally step 320 of dynamic culture conditions. Step 310 may comprise seeding at least one IPHB with at least one cell type. The IPHB may comprise a 3D continuous polymeric matrix with a network of microporous cavities. The cavities may comprise one or more channels, pores, and / or chambers. Step 320 may comprise exposing cells to dynamic culture conditions. Dynamic culture conditions 320 may be configured to achieve nutrient and / or waste exchange. Dynamic culture conditions 320 may comprise exposing at least one cell to a culture media via perfusion and / or static solution. The culture media may comprise one or more nutrients. Dynamic culture conditions 320 may comprise varying a culture media’s nutrient type and / or concentration. Dynamic culture conditions 320 may comprise perfusing at least one block with culture media. In certain embodiments, one or more cavities may be configured as pathway(s) for nutrient delivery, gas exchange, and / or waste removal.

[0104] In some embodiments of step 310, at least one cell may be derived from a human, a subject, a biopsy, or a combination thereof. The biopsy could be from the subject or otherwise. The subject could be a human or a nonhuman. Multiple specimens could be from the same and / or different sources; examples include multiple specimens from the same biopsy, different biopsies from the same subject (e.g. including differing in the location and / or time biopsied), and / or different subjects. One or more subjects may be mammals, animals, birds, reptiles, amphibians, and / or fish. At least one cell could be derived from one or more tissues and / or organs. Additional permutations will be readily apparent to those of ordinary skill in the art.

[0105] Certain embodiments may comprise additional or different steps. For example, step 320 is optional. By way of further example, method 300 may comprise step 330 of at least one personalized model and / or step 340 of at least one analysis. Step 330 may comprise cultivating cells derived from one or more subjects into one or more personalized models. Step 340 may comprise subjecting cells (e.g., the personalized model(s)) to one or more analyses to guide diagnosis and / or therapy, which could be tailored for the subjects). Step 340 may comprise exposing one or more of the cells to at least one experimental variable, which could be across one or more geographic, temporal, and / or other differential(s) and / or gradients). An experimental variable may comprise at least one drug, thermal energy, radiation, or any combination thereof. A drug may comprise at least one medicine, medication, pharmaceutical, biologic, immunologic, immunotherapy, chemotherapy, radiotherapy, radiotracer, tracer, stain, diagnostic agent, theranostic agent, therapeutic agent, and / or treatment agent. Step 340 may comprise evaluating, measuring, and / or observing the effect of at least one experimental variable on the senescence, growth, and / or characteristic(s) of the cells (e.g., seeded, cultivated, primary, tumor, metastatic, tissue, and / or organ cells).

[0106] All steps may be incorporated in any combination and in any order, including repeating step(s). By way of example, certain method embodiments may comprise seeding 310, dynamic culture conditions 320, and analysis 340 (not necessarily including personalized model 330). In some embodiments, personalized model 330 could be after or subsequent to analysis 340. Some embodiments might not comprise dynamic culture conditions 320. Some embodiments may comprise cyclical iterations, in any order, such as multiple instances of seeding 310, dynamic culture conditions 320, personalized model 330, and / or analysis 340. Further, the one or more blocks underlying step 310 (and / or being seeded) could be altered before, during, and / or after other steps (e.g. seeding 310, dynamic culture conditions 320, personalized model 330, and / or analysis 340). Such embodiments may include changing the configuration of existing blocks, removing blocks, and / or adding blocks.

[0107] Throughout embodiments of systems, methods, and blocks herein, cells may comprise one or more adipose derived MSCs, HEK 293 cells, HeLa cells, Henrietta Lacks cells, CHO cells, Chinese hamster ovary cells, Wharton jelly cells, bone marrow derived MSCs, Panc-1 cells, ASPC-1 cells, BcPc-3 cells, pancreatic cancer cells, human pancreatic cancer cells, endothelial cells, keratinocytes, keratocytes, CD4+ T cells, CD8+ T cells, IPSCs, PBMC cells, peripheral blood mononuclear cells, hepatocytes, porcine hepatocytes, muscle derived MSCs, equine muscle derived MSCs, MDA-MCB-231 cells, breast cancer cells, human breast cancer cells, triple negative breast cancer cells, bone marrow aspirate, human bone marrow aspirate, SNU-398 cells, HepG2 cells, liver cancer cells, human liver cancer cells, SHSY5Y cells, neuroblastoma cells, human neuroblastoma cells, murine neuroblastoma cells, N2a cells, mouse neuroblastoma cells, neuroprogenitor cells, Schwann cells, macrophages, dorsal root ganglia, murine dorsal root ganglia, myocytes, bovine myocytes, chondrocytes, porcine chondrocytes, osteocytes, porcine osteocytes, UB-OC2 cells, organ of Corti cells, murine organ of Corti cells, cochlear cells, prostate PDX cells, PC3 cells, prostate cancer cells, human prostate cancer cells, A549 cells, lung carcinoma cells, human lung carcinoma cells, DMS53 cells, H82 cells, epithelial like lung carcinoma cells, NIH-H1703 cells, squamous lung cancer cells, A673 cells, muscle cancer cells, soft tissue cancer cells, bone cancer cells, sarcoma cells, Ewing sarcoma cells, human Ewing sarcoma cells, MDCK cells, Madin-Darby canine kidney cells, planaria slurry, snail slurry, kidney slurry, murine kidney slurry, kidney explant, murine kidney explant, Vero cells, monkey Vero cells, epithelial cells, E. coli, S. aureus, trabecular meshwork cells, renal cells, human renal cells, C2C12 cells, myoblasts, myoblast cells, murine myoblast cells, MLO-A5 cells, IDG-SW3 cells, osteoblast cells, osteocyte-like cells, murine osteoblast cells, murine osteocyte-like cells, fibroblast cells, fibroblasts, human fibroblasts, porcine fibroblasts, avian fibroblasts, fish fibroblasts, murine fibroblasts, American Type Culture Collection (ATCC) fibroblasts, CRL fibroblasts, CRL-4061 fibroblasts, hTERT gingival fibroblasts, CRL-4058 fibroblasts, hTERT lung fibroblasts, CRL-2522 fibroblasts, BJ fibroblast cells, NIH / 3T3 cells, embryonic fibroblast cells, murine embryonic fibroblast cells, or any combination thereof.

[0108] Persons having ordinary skill in the art will readily appreciate that the various features of embodiments herein, whether discussed or mentioned or disclosed in a context of a system, method, or block, may be practiced in embodiments of other / different blocks, methods, and / or systems, and vice versa, all within the present scope.

[0109] As explained above and reiterated below, the present disclosure includes, without limitation, the following example implementations.

[0110] Clause 1. A modular cell culture system comprising: a plurality of IPHBs, wherein each of the IPHBs comprises a biocompatible hydrogel matrix with an interconnected network of microchannels, and wherein each of the IPHBs are configured to interconnect horizontally and / or vertically to one or more additional IPHBs to form a customizable and expandable culture environment; and the microchannels are configured to provide pathways for nutrient delivery, gas exchange, and waste removal

[0111] Clause 2. The system of clause 1 , wherein the hydrogel matrix of each respective IPHB is configured to mimic the ECM of native tissues of interest, supporting natural cell adhesion, growth, and differentiation.

[0112] Clause 3. The system of clause 2, wherein the hydrogel matrix of each respective IPHB is configured to be oxygen-permeable and swellable based on the salinity of the perfused liquid.

[0113] Clause 4. The system of clause 1, wherein the microchannels of each respective IPHB are configured to form a pseudo-vasculature network facilitating the efficient transport of nutrients and removal of waste products.

[0114] Clause 5. The system of clause 4, wherein each respective IPHB is configured to support both static and perfusion-based cultures, allowing for controlled delivery of nutrients, gases, and mechanical stimuli.

[0115] Clause 6. The system of clause 1 , wherein each respective IPHB is configured to enable the co-culture of multiple cell types within the same system to facilitate the formation of complex tissue structures and organ components.

[0116] Clause 7. The system of clause 1, wherein each respective IPHB is configured to be enzymatically dissolved to release cells for downstream applications, preserving cell viability and functionality; wherein the biocompatible hydrogel matrix is enzymatically dissolvable. Clause 8. The system of clause 1, wherein each respective IPHB is configured for histological and immunohistochemical analysis to allow for detailed examination of tissue architecture and cellular interactions.

[0117] Clause 9. The system of clause 1, wherein each respective IPHB is configured to reduce media and plasticware consumption compared to traditional cell culture methods, lowering overall production costs.

[0118] Clause 10. A system for personalized medicine, comprising: a plurality of IPHBs configured to support the co-culture of patient-specific cells, wherein the plurality of IPHBs are configured to provide the capability to form patient-specific tissue models for drug testing and therapy optimization.

[0119] Clause 11. The system of clause 10, wherein the plurality of IPHBs are configured to allow for the comparison of healthy and diseased tissues to identify specific drug targets and optimize treatment protocols for individual patients.

[0120] Clause 12. A method for producing organized tissue structures and functional organ components, comprising: seeding cells into the microchannels of a plurality of IPHBs; interconnecting the plurality of IPHBs horizontally and / or vertically to expand the culture environment; and providing nutrients, gases, and mechanical stimuli through static and / or perfusion-based cultures to promote cell growth and tissue organization.

[0121] Clause 13. The method of clause 12, wherein each of the IPHBS include a respective hydrogel matrix being configured to support the formation of a pseudo-vasculature network within the tissue constructs.

[0122] Clause 14. The method of clause 13, wherein each respective hydrogel matrix is configured to be oxygen-permeable and swellable, enhancing cell viability and function.

[0123] Clause 15. The method of clauses 13-14, further comprising enzymatically dissolving the hydrogel matrix to release cells for downstream applications.

[0124] Clause 16. The method of clause 12, further comprising a step of co-culturing multiple cell types within the plurality of IPHBs to facilitate the development of complex tissue structures and organ components.

[0125] Clause 17. The method of clause 12, wherein at least one of the plurality of IPHBs is configured to enable the production of high-density cell cultures and high yields of biologies, improving cost-effectiveness and scalability. Clause 18. The method of clause 12, wherein the modular and expandable nature of the plurality of IPHBs is configured to support both small-scale research and large-scale industrial applications.

[0126] Clause 19. A method for enhancing drug discovery and screening, comprising: using at least one IPHB to create high-throughput, physiologically relevant models for preclinical studies; and assessing drug efficacy and toxicity in the tissue models formed within the at least one IPHB.

[0127] Clause 20. The method of clause 19, wherein the at least one IPHB is configured to enable detailed analysis of cellular responses and interactions, providing comprehensive insights into experimental outcomes.

[0128] These and other modifications and variations may be practiced by those of ordinary skill in the art without departing from the spirit and scope of the claims. In addition, it should be understood that aspects of the various embodiments may be interchanged in whole or in part. Furthermore, those of ordinary skill in the art will appreciate that the foregoing description is by way of example only, and it is not intended to limit the claims. Therefore, the spirit and scope of the appended claims should not be limited to the exemplary description of the versions contained herein.

Claims

CLAIMSTHAT WHICH IS CLAIMED:

1. A modular cell culture system, comprising a plurality of interconnecting porous hydrogel blocks, wherein at least one block comprises a three-dimensional continuous polymeric matrix with a network of microporous cavities and is configured to interconnect with at least one other block, wherein one or more cavities are configured as pathways for nutrient delivery, gas exchange, waste removal, or a combination thereof.

2. The system of claim 1 , wherein one or more blocks are configured to mimic an extracellular matrix.

3. The system of claim 2, further configured to match at least one physical property of the extracellular matrix.

4. The system of claim 3, wherein one or more physical properties are selected from the group consisting of: porosity, stiffness, hardness, capacitance, elasticity, elastic modulus, density, conductivity, thermal conductivity, electric conductivity, electric charge, solubility, hydrophilicity, lipophilicity, or a combination thereof.

5. The system of claim 1 , wherein at least one block is oxygen permeable.

6. The system of claim 5, wherein at least one block is oxygen permeable based on the salinity of at least one perfused fluid.

7. The system of claim 1, wherein at least one block is swellable.

8. The system of claim 7, wherein at least one block is swellable based on the salinity of at least one perfused fluid.

9. The system of claim 1, wherein a plurality of pathways mimic pseudo- vasculature.

10. The system of claim 10, configured to permit exposure of one or more cavities to at least one nutrient, gas, mechanical stimulus, static solution, perfusion, or any combination thereof.

11. The system of claim 1, wherein the cavities comprise one or more channels, one or more chambers, one or more pores, or a combination thereof.

12. The system of claim 1, wherein at least one block is configured to be enzymatically dissolvable.

13. The system of claim 1, wherein at least one block is configured for histological or immunohistochemical study or a combination thereof.

14. The system of claim 1, further configured to cultivate one or more cell types when seeded with at least one cell of each type.

15. The system of claim 14, configured to permit co-cultivation of a plurality of cell types within at least one block.

16. The system of claim 14, wherein one or more cells are derived from a specimen, a human, a subject, a biopsy, or a combination thereof.

17. The system of claim 1, wherein at least one cavity is configured for communication between two or more interconnected blocks in one or more vertical, horizontal, or orthogonal axes relative to gravity or any combination thereof.

18. A modular cell culture method, comprising seeding at least one interconnecting porous hydrogel block with at least one cell type, wherein at least one block comprises a three-dimensional continuous polymeric matrix with a network of microporous cavities and is configured to interconnect with at least one other block,further wherein one or more cavities are configured as pathways for nutrient delivery, gas exchange, waste removal, or a combination thereof.

19. The method of claim 18, wherein at least one cell is isolated from a subject.

20. The method of claim 19, further comprising cultivating the cells into a personalized model and subjecting the model to analysis to guide diagnosis, therapy, or a combination thereof tailored for the subject.

21. A modular cell culture block, comprising a three-dimensional continuous polymeric matrix with a network of microporous cavities, configured to interconnect with at least one other block, wherein one or more cavities are configured as pathways for nutrient delivery, gas exchange, waste removal, or a combination thereof.

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