Method of cryopreserving selected sperm cells
DC CAFCFirst Claim
1. A method of cryopreserving sex-selected sperm cells, comprising:
- a. obtaining sperm cells from a species of a non-human male mammal;
b. sorting said sperm cells, without the presence of protective compounds in seminal plasma, and based upon sex-type to provide a collection of sex-selected sperm cells obtained using flow cytometry or fluorescence-activated cell sorting;
c. cooling said sex-selected sperm cells;
d. suspending said sex-selected sperm cells in an extender to provide a concentration of sperm cells of about 5 million per milliliter of extender to about 10 million per milliliter of extender; and
e. freezing said sex-selected sperm cells in said extender.
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Abstract
The present invention provides a method of cryopreserving sperm that have been selected for a specific characteristic. In a preferred embodiment, the method is employed to freeze sex-selected sperm. Although the cryopreservation method of the invention can be used to freeze sperm selected by any number of selection methods, selection using flow cytometry is preferred. The present invention also provides a frozen sperm sample that has been selected for a particular characteristic, such as sex-type. In preferred embodiments, the frozen sperm sample includes mammalian sperm, such as, for example, human, bovine, equine, porcine, ovine, elk, or bison sperm. The frozen selected sperm sample can be used in a variety of applications. In particular, the sample can be thawed and used for fertilization. Accordingly, the invention also includes a method of using the frozen selected sperm sample for artificial insemination or in vitro fertilization.
367 Citations
28 Claims
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1. A method of cryopreserving sex-selected sperm cells, comprising:
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a. obtaining sperm cells from a species of a non-human male mammal; b. sorting said sperm cells, without the presence of protective compounds in seminal plasma, and based upon sex-type to provide a collection of sex-selected sperm cells obtained using flow cytometry or fluorescence-activated cell sorting; c. cooling said sex-selected sperm cells; d. suspending said sex-selected sperm cells in an extender to provide a concentration of sperm cells of about 5 million per milliliter of extender to about 10 million per milliliter of extender; and e. freezing said sex-selected sperm cells in said extender. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28)
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Specification