Three-dimensional bioreactors
Inventors
Ling, Jian • Wellinghoff, Stephen T. • RUBAL, Michael Joseph
Assignees
Southwest Research Institute SwRI
Publication Number
US-11447731-B2
Publication Date
2022-09-20
Expiration Date
2039-09-24
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Abstract
The present invention relates to the design, fabrication and applications of three-dimensional (3D) bioreactor for cell expansion and cell secreted substance production. The bioreactors have relatively low levels of potentially cytotoxic compounds, can be coated with substituted or unsubstituted poly(p-xylene) type coatings and can also be separately formed from liquid crystal photopolymerizable monomers.
Core Innovation
The invention concerns a three-dimensional (3D) bioreactor designed for cell expansion and production of cell-secreted substances. It is distinguished by a structure comprising a plurality of interconnected voids for cell growth, each defined by precise, non-random void volumes and pore openings, with more than 90% of voids and pores having their volumes and diameters, respectively, within ±10% tolerance. The bioreactor is fabricated from materials such as biocompatible polymers or photopolymerizable liquid crystalline monomers and is optionally coated with substituted or unsubstituted poly(p-xylene), further functionalized with polydopamine, to enable additional biofunctional coatings.
The problem being addressed is the inefficiency, high cost, and contamination risk inherent in current cell expansion methods, particularly for T-cell and stem cell therapies. Existing approaches rely on magnetic beads and complex multi-stage open systems, which increase manual intervention, contamination risk, and expense, making them less suitable for large-scale and cost-effective clinical manufacturing. Moreover, conventional 2D culture methods and existing 3D devices suffer from limited scalability, inadequate process control, high shear stresses, and processing complexities.
The 3D bioreactor overcomes these limitations through regular, tunable geometry that offers a continuous, high surface-to-volume ratio suitable for cell culturing, improved flow characteristics, and lower shear stress for the cells. Its design supports scalable manufacturing by additive techniques and minimizes cytotoxic leaching through material selection, post-processing with UV/heat/vacuum, and surface coating. The modular coating approach, including polydopamine layering over parylene-type coatings, facilitates the immobilization of proteins, antibodies, or extracellular matrix for both non-adherent and adherent cell expansion applications.
Claims Coverage
There are two independent claims that define the main inventive features of this patent.
3D bioreactor with precise voids and pore openings, poly(p-xylylene) coating, and polydopamine layer
A three-dimensional bioreactor for cell growth comprising: - A plurality of voids providing surface area for cell expansion, where each void has a diameter D and each pore opening between voids has a diameter d, and D > d. - At least 90% of the voids have a selected void volume (V) that does not vary by more than ±10%. - At least 90% of the pore openings have a value of d that does not vary by more than ±10%. - The bioreactor is coated with a substituted or unsubstituted poly(p-xylylene). - Polydopamine is applied to the poly(p-xylylene) coating, and the polydopamine is capable of binding an additional layer or layers for cell culturing.
Method for forming a 3D bioreactor by additive manufacturing and functional coatings
A method of forming a 3D bioreactor comprising: 1. Forming the 3D bioreactor by additive manufacturing so that the bioreactor includes a plurality of voids with surface area for cell expansion, each void having a diameter D, and pore openings of diameter d, with D > d, and at least 90% of voids and pores within ±10% tolerance in size. 2. Coating the bioreactor with substituted or unsubstituted poly(p-xylylene). 3. Applying polydopamine to the poly(p-xylylene) coating, with the polydopamine capable of binding additional layer(s) for cell culturing.
These inventive features center on a precisely engineered 3D bioreactor architecture with tight dimensional tolerances, biologically relevant surface coatings, and scalable fabrication via additive manufacturing combined with functionalization strategies for enhanced cell culture performance.
Stated Advantages
The 3D bioreactor provides a high surface-to-volume ratio, enabling large cell expansion within a small footprint.
The continuous, interconnected surface allows cells to migrate freely, in contrast to fragmented surfaces of other reactors.
Uniform and low shear stress within the reactor reduces the risk of shear-induced cell differentiation.
The design supports uniform nutrient and oxygen flow, minimizing gradients and improving cell culture conditions.
Precise, non-random geometry enables reproducible manufacturing and predictable bioreactor performance.
Functional coatings, including poly(p-xylene) and polydopamine, reduce cytotoxic leaching and support further biofunctionalization for diverse cell types.
The bead-free, closed-loop system for T-cell expansion simplifies the process, reduces contamination risk, and lowers cost.
Additive manufacturing techniques provide scalable and cost-effective fabrication.
Documented Applications
Expansion of T-cells for immunotherapy, such as CAR T-cell therapy.
Expansion of stem cells, including mesenchymal and adipose-derived stem cells, for regenerative medicine.
Production of cell-secreted substances.
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