System and process for continuous addition of cryoprotectant
Inventors
Busa, Domenic • Botts, Michael • Klas, Gary • Parker, Johnathan
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11889830-B2
Publication Date
2024-02-06
Expiration Date
Abstract
An integrated system and method for preparing sperm cells to improve their survivability during cryopreservation are described herein. The system features a vessel, a controlled dispenser, and a dispense tube. The dispense tube has a first end fluidly connected to the dispenser and a second end disposed inside the vessel. The second end can be submerged in the sperm cell fluid. The controlled dispenser may be a syringe pump that includes a syringe for containing a cryoprotectant and a pushing mechanism for displacing the syringe. The syringe pump is configured to discharge the cryoprotectant through the dispense tube and into the vessel, thereby dispensing the cryoprotectant into the sperm cell fluid. Mixing of the fluids is achieved using a shaker table agitation.
Core Innovation
The cryopreservation system continuously dispenses a cryoprotectant into a vessel containing biological specimen, including sperm cells. A controlled dispenser dispenses cryoprotectant through a dispense tube having a first end fluidly connected to a discharge port and a second end disposed inside the vessel, where the second end contacts an internal surface of the vessel or is submerged in material contained within the vessel to prevent droplet formation and dripped addition.
The system uses controller-actuated dispensing of cryoprotectant through the dispense tube into the vessel in a manner or flow rate such that the cryoprotectant is dispensed without being dripped into the vessel. The dispenser provides programmable flow-rate control including continuous or pulsed/interrupted dispensing modes, and a mixer provides agitation on a shaker-table while dispensing to reduce cell stress and reduce local concentration spikes.
The description further supports multi-dispense and multi-fluid arrangements, including additional cryoprotectant or gas addition, and optional temperature control. Documented results indicate increased cell survivability relative to step-wise bulk additions and increased post-freeze/thaw insemination dose relative to step-wise bulk additions.
Claims Coverage
The provided material includes three independent claims, directed to a cryoprotectant dispensing system, a cryopreservation system with a syringe pump, and a method. Across these independent claims, the main inventive aspects are the dispense tube second-end contacting/submerging arrangement to prevent droplet formation and dripping, controller-controlled dispensing at a desired manner or flow rate, and a syringe-pump controlled delivery architecture in one claim set.
Droplet prevention by tube second-end contact or submerging
A dispense tube has a second end disposed inside the vessel so that the second end contacts an internal surface of the vessel or is submerged in material contained within the vessel, thereby preventing droplet formation and dispensing the cryoprotectant without being dripped into the vessel.
Controller-actuated dispensing of cryoprotectant at a manner or flow rate
A controller comprising a processor and a memory actuates a controlled dispenser to dispense a cryoprotectant fluid through the dispense tube into the vessel in a manner or flow rate such that droplet formation is prevented and the cryoprotectant is dispensed without being dripped into the vessel.
Syringe pump with motor-controlled pusher step size for desired flow rate
A syringe pump includes a syringe and a pushing mechanism with a moveable pusher block, a motor, and a motor controller including a processor and memory that stores instructions to cause the motor to move the pusher block at a desired step size, such that the cryoprotectant is added to the biological specimen at a desired flow rate.
Continuous or pulsed cryoprotectant addition via controlled flow rate
The system dispenses the cryoprotectant into the biological specimen at a flow rate such that the addition is continuous or alternatively pulsed, depending on the specified delivery mode.
Mixer providing agitation while preventing adherence or local concentration spikes
A mixer mixes the cryoprotectant and biological specimen in the vessel while preventing the specimen from adhering to the vessel internal surface.
Multi-dispense configuration dispensing same or different fluids
The system includes one or more additional controlled dispensers connected by a dispense tube to a vessel, where the dispensers dispense the same or different fluids into the vessel.
Method dispensing cryoprotectant without dripped addition via tube contact
A method for dispensing cryoprotectant to a biological specimen by fluidly coupling cryoprotectant to a controlled dispenser, adding the biological specimen into a vessel, connecting the controlled dispenser to the vessel via a dispense tube with a second end disposed inside and contacting the biological specimen or an internal surface, and dispensing the cryoprotectant at a desired flow rate such that droplet formation is prevented and the cryoprotectant is dispensed without being dripped into the vessel.
The independent claims collectively cover a dispense tube arranged so the second end contacts the vessel internal surface or the specimen to prevent droplet formation and dripped addition, controller-controlled dispensing of cryoprotectant into the vessel at a desired manner or flow rate, a syringe-pump architecture with a motor controller moving a pusher block at a desired step size to achieve a desired flow rate, and dependent refinements including mixing/agitation and multi-dispense arrangements for same or different fluids.
Stated Advantages
Droplet formation is prevented and the cryoprotectant is dispensed without being dripped into the vessel.
Reduced cell stress and reduced local concentration spikes are stated in the described system.
Increased cell survivability is stated relative to step-wise bulk additions.
Increased post-freeze/thaw insemination dose is stated relative to step-wise bulk additions.
Documented Applications
Cryopreservation of sperm, including dispensing a cryoprotectant into a vessel containing sperm cells.
Post-freeze/thaw insemination use, associated with increased insemination dose in the described example.
Interested in licensing this patent?