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Publication Number
US-10961501-B2
Publication Date
2021-03-30
Expiration Date
Abstract
A system for seeding therapeutic active ingredients (TAIs) onto a porous scaffold includes a first chamber for accommodating the scaffold, a TAI storage device for storing TAIs, at least one second chamber for storing TAI media, and a gas inlet for receiving gas from a compressed source. A flow circuit is coupled to the first chamber, the TAI storage device, the second chamber and the gas inlet for delivering the TAIs, the TAI media and the gas to the scaffold. A pump pumps at least one of the TAIs, the TAI media and the gas in the flow circuit. The system also includes a processor that regulates the delivery of the TAIs, the TAI media and the gas to the scaffold via a plurality of valves.
Core Innovation
The disclosed invention provides an automated bioreactor system and associated methods for seeding and culturing therapeutic active ingredients (TAIs) in a porous scaffold by combining liquid and gas pressure. A first chamber holds the porous scaffold while TAIs, TAI media, and gas are delivered through a flow circuit. The system includes a TAI storage device, a pump, a gas inlet from a compressed sterile gas source, and a processor that controls a plurality of valves to coordinate delivery and waste flow.
The method includes a multi-phase seeding process in which the porous scaffold surface is wetted and coated with a thin layer to facilitate wicking of a cell solution and to result in a uniform seeded scaffold. Forced-media pre-wet fills pores, and gas purging uses gas flow and pulsed pressurized gas bursts to remove excess media while leaving a thin wetting layer for capillary wicking and uniform coverage. A scaffold turner/rotary holder is used to rotate the scaffold to counter gravity-driven bias and improve uniform adhesion.
The invention further describes a culturing phase in which TAI media is refreshed during culturing over weeks. The processor coordinates periodic valve switching and pumping, while scaffold rotational orientation changes are used to support uniform growth and attachment within the porous scaffold. The document reports homogenous delivery/attachment of fluorescently labeled ATIs in large porous scaffolds, and describes alternative configurations such as optional waste valve and manual operation.
Claims Coverage
The provided claim set includes one independent method claim, with dependent claims refining the delivery setup and control strategy. The independent claim centers on wetting and coating a porous scaffold with a thin layer using liquid and gas pressure to facilitate wicking and achieve a uniform seeded scaffold.
Wetting and coating with thin layer to facilitate wicking for uniform seeding
Using liquid and gas pressure to wet and coat the surface of the porous scaffold with a thin layer to facilitate wicking of a cell solution, resulting in a uniform seeded scaffold.
Chambered delivery with processor-controlled valves
Accommodating the porous scaffold in a first chamber and delivering TAIs, TAI media, and gas via a flow circuit from a TAI storage device using a pump, with a processor regulating delivery through a plurality of valves.
Valve-specific regulation of media, gas, TAIs, chamber delivery, and waste
The plurality of valves includes a media valve, a gas valve, a TAI valve, a first chamber valve, and a waste valve to regulate respective deliveries and waste flow.
Pulsed pressurized gas bursts by alternating valve open/closed states
The processor regulates pulsing pressurized gas bursts through the scaffold by repeatedly alternating the gas valve, the first chamber valve, and the waste valve between open and closed positions while keeping remaining valves closed.
Pressurization phase prior to releasing gas burst
The processor switches the gas valve to an open position and the first chamber valve to a closed position to generate pressurized gas prior to releasing a pressurized gas burst.
Release phase into the first chamber
The processor switches the gas valve to a closed position and the first chamber valve to an open position to release the pressurized gas burst into the first chamber.
Overall, the claim coverage maps a uniform seeding mechanism based on thin-layer wetting via liquid and gas pressure, and then refines it into a chambered delivery architecture with processor-controlled valves that implement a pulsed pressurized gas burst sequence through alternating gas, first-chamber, and waste valve states, including explicit pressurization and release valve configurations.
Stated Advantages
Uniform seeded scaffold by wetting/coating with a thin layer that facilitates wicking of a cell solution.
Homogenous delivery/attachment of fluorescently labeled ATIs in large porous scaffolds.
Improved uniform adhesion by rotating the scaffold to counter gravity-driven bias.
Uniform growth and attachment during culturing supported by refreshing TAI media and changing rotational orientation.
Documented Applications
Seeding and culturing therapeutic active ingredients (TAIs) in porous scaffolds in an automated bioreactor system.
Delivering and achieving attachment of fluorescently labeled ATIs in large porous scaffolds.
Multi-phase workflow including forced-media pre-wet, gas purging with pulsed pressurized gas bursts, and media refreshing during culturing.
Alternative operation/configurations including optional waste valve and manual operation.
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