Multiple sexed embryo production system for mammals using low numbers of spermatozoa
AssigneeXy, LLC
InventorsGeorge Seidel, Lisa Herickhoff, John Schenk
Life of this patent
Estimating the term…
Abstract
Improved insemination systems particularly adapted to use for sex-selected sperm sorting include systems which achieve superovulation and then multiple embryo production with sexed embryos. These systems combine with other techniques, including techniques for enhanced sheath fluid and other strategies which minimize stress on the sperm cells, and, potentially, a 2.9 percent sodium citrate sheath solution for bovine species and a hepes bovine gamete media for equine species. Improved collection systems and techniques for the process are described so that commercial application of sperms samples as well as the resulting animals may be achieved.
Claims143
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells which contains about 2.9% sodium citrate;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells which contains a hepes buffered medium;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed and which comprises a citrate collector fluid containing about six percent egg yolk
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed and which comprises a test tube having the physical characteristics of a stream-matched container.
An improved flow cytometer system for isolating desired cells as described in claim 8 wherein said cell source comprises cells which are mechanically delicate.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a chemically coordinated sheath fluid source which creates a sheath fluid environment for said cells which is selected to be coordinated with both a pre-sort and a post-sort cell fluid environment;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed.
An improved flow cytometer system for isolating desired cells as described in claim 22 wherein said cell source comprises cells which are hyper-responsive to a chemical composition in a sheath fluid environment.
An improved flow cytometer system for isolating desired cells as described in claim 39 wherein said cell source comprises bovine sperm cells.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells which comprises a solution containing about 2.9% sodium citrate;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed.
An improved flow cytometer system for isolating desired cells as described in claim 40 wherein said collector is used to provide a sperm for artificial insemination.
An improved flow cytometer system for isolating desired cells as described in claim 40 wherein said collector is used to provide a low dose of sperm for artificial insemination.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells which comprises a solution containing a hepes buffered medium;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed.
An improved flow cytometer system for isolating desired cells as described in claim 45 wherein said collector is used to provide a low dose of sperm for artificial insemination.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies bovine sperm cells to be analyzed by the flow cytometer,
- b. a chemically coordinated sheath fluid source which creates a sheath fluid environment for said cells which is contains about 2.9% sodium citrate;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- g. a collector into which cells having a desired characteristic which contains a citrate collection fluid comprising about six percent egg yolk and which is used to provide a dosage of less than about five hundred thousand sperm;
- and wherein said nozzle, oscillator, cell sensing system, and sorter discrimination system are part of a flow cytometer system which sorts said cells to be analyzed at a rate of at least about five hundred sorts per second and operates at a pressure of at least about icy pounds per square inch.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies equine sperm cells to be analyzed by the flow cytometer,
- b. a chemically coordinated sheath fluid source which creates a sheath fluid environment for said cells which is contains a hepes buffered medium;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- g. a collector into which cells having a desired characteristic which contains a collection fluid comprising a hepes buffered medium and which is used to provide a dosage of less than about ten million sperm;
- and wherein said nozzle, oscillator, cell sensing system, and sorter discrimination system are part of a flow cytometer system which sorts said cells to be analyzed at a rate of at least about five hundred sorts per second and operates at a pressure of at least about fifty pounds per square inch.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a means for minimizing the changes between a sheath fluid environment for said cells and both a pre-sort and a post-sort cell fluid environment;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector into which cells having a desired characteristic are placed which comprises a chemically coordinated collector fluid sheath fluid source which creates a collector fluid environment for said cells which is selected to be coordinated with a prior cell fluid environment.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acres to sort cells having a desired characteristic;
- and
- g. a collector which comprises a cushioning element.
An improved flow cytometer system for isolating desired cells as described in claim 59 wherein said container comprises a test tube having the physical characteristics of a stream-matched container.
An improved flow cytometer system for isolating desired cells comprising:
- a. a cell source which supplies cells to be analyzed by the flow cytometer,
- b. a sheath fluid source which creates a sheath fluid environment for said cells;
- c. a nozzle through which said cells pass while subjected to said sheath fluid environment;
- d. an oscillator which acts upon said sheath fluid as it passes through said nozzle;
- e. a cell sensing system which responds to said cells;
- f. a sorter discrimination system which acts to sort cells having a desired characteristic;
- and
- g. a collector which comprises a means for avoiding impact between said cells and said collector.
A method of sorting cells comprising the steps of:
- a. establishing a cell source which supplies cells to be sorted;
- b. chemically coordinating a sheath fluid to create a sheath fluid environment for said cells which is coordinated with both a pre-sort and a post-sort cell fluid environment;
- c. sensing a property of said cells;
- d. discriminating between cells having a desired characteristic;
- and
- e. collecting cells having the desired characteristic.
A method of sorting cells comprised the steps of:
- a. establishing a cell source which supplies cells to be sorted;
- b. establishing a sheath fluid to create a sheath fluid environment for said cells;
- c. sensing a property of said cells;
- d. discriminating between cells having a desired characteristic;
- e. collecting cells having the desired characteristic in a collector fluid;
- and
- f. chemically coordinating said collector fluid to create an ending collector fluid environment for said cells which is coordinated with a pre-sort fluid environment.
A method of sorting cells comprising the steps of:
- a establishing a cell source which supplies cells to be sorted;
- b. establishing a sheath fluid to create a sheath fluid environment for said cells;
- c. sensing a property of said cells;
- d. discriminating between cells having a desired characteristic;
- e. collecting cells having the desired characteristic comprising the step of cushioning said cells from impact with a collection container.
A method of producing multiple, sexed embryos from a female mammal comprising:
- a. creating superovulation in said mammal to create at least two eggs comprising the step of using an ovulatory pharmaceutical to cause multiple eggs to be produced;
- b. determining a sex of a sperm cell of a male mammal;
- c. sorting according to said sex of said sperm cells;
- d. inserting at least a portion of said sorted sperm cells into said uterus after an onset of estrus;
- and
- e. fertilizing a plurality of said eggs to produce at least one animal of the desired sex.
A method of sorting cells comprising:
- a. establishing a cell source which supplies cells to be sorted;
- b. chemically coordinating a sheath fluid to create a sheath fluid environment for said cells which is coordinated with both a pre-sort and a post-sort cell fluid environment;
- c. sensing a property of said cells;
- d. discriminating between cells having a desired characteristic;
- and
- e. collecting cells having the desired characteristic.
Dependencies are inferred from the claim text.