Sex selected equine embryo production system

• US 20110078803A1
• Filed: 08/24/2010
• Published: 03/31/2011
• Est. Priority Date: 08/01/2002
• Status: Active Grant

• Current Assignee: XY, LLC (Inguran LLC)
• Original Assignee: XY, LLC (Inguran LLC)
• Inventors: Schenk, John L., Seidel, George E., Suh, Tae Kwang
• Granted Patent: US 8,497,063 B2
• Time in Patent Office: Days
• US Class Current: 800/8
• CPC Class Codes: A01K 2227/10 Mammal
• CPC Class Codes: A01K 2227/101 Bovine
• CPC Class Codes: A01K 67/027 New or modified breeds of vertebrates
• CPC Class Codes: A61D 19/00 Instruments or methods for reproduction or fertilisation
• CPC Class Codes: C12N 2517/10 Conditioning of cells for in vitro fecondation or nuclear transfer
• CPC Class Codes: C12N 5/061 Sperm cells, spermatogonia
• CPC Class Codes: C12N 5/0612 sorting of gametes, e.g. according to sex or motility
• CPC Class Codes: Y10T 442/2115 At least one coating or impregnation functions to fix pigments or particles on the surface of a coating or impregnation

Abstract

A sex selected equine embryo production system for the production of sex selected equine embryos and offspring by fertilization of oocytes with sex selected sperm by intracytoplasmic sperm injection.

Claims

1-64. -64. (canceled)

65. A method of producing an equine animal, comprising the steps of:

a) obtaining a plurality of equine sperm cells;

b) determining a sex of said plurality of equine sperm cells;

c) sorting said plurality of equine sperm cells based upon said sex;

d) obtaining an equine oocyte;

e) intracytoplasmically injecting said equine oocyte with one of said plurality of equine sperm cells of a selected sex; and

f) producing an equine embryo of said selected sex.

66. The method of producing an equine animal as described in claim 65, further comprising the step of obtaining a recipient animal capable of receiving said equine embryo of said selected sex.

67. The method of producing an equine animal as described in claim 66, further comprising the step of transferring said equine embryo of said selected sex to said recipient animal.

68. The method of producing an equine animal as described in claim 67, further comprising the step of gestating said equine embryo within said recipient animal for a period of time.

69. The method of producing an equine animal as described in claim 65, further comprising the step of establishing discrete populations of equine sperm cells of same said sex.

70. The method of producing an equine animal as described in claim 69, further comprising the step of freezing said equine sperm cells of same said sex.

71. The method of producing an equine animal as described in claim 70, further comprising the step of thawing said equine sperm cells of same said sex.

72. The method of producing an equine animal as described in claim 71, further comprising the step of washing said equine sperm cells of same said sex.

73. The method of producing an equine animal as described in claim 72, wherein said step of washing comprises washing said equine sperm cells of same said sex in FCDM.

74. The method of producing an equine animal as described in claim 72, further comprising the step of swimming up said equine sperm cells of same said sex prior to said step of washing.

75. The method of producing an equine animal as described in claim 74, wherein said step of swimming up comprises the step of swimming up of said equine sperm cells of same sex in CDM containing about 2 mM caffeine and heparin.

76. The method of producing an equine animal as described in claim 65, further comprising the step of utilizing a piezo injection system for injecting one of said plurality of equine sperm cells Of said selected sex into said equine oocyte.

77. The method of producing an equine animal as described in claim 76, further comprising the step of providing a sperm-injection pipette having an external diameter of about eight to ten micrometers.

78. The method of producing an equine animal as described in claim 65, further comprising the step of providing a flow cytometer which performs said steps of determining a sex of said plurality of equine sperm cells based on said sex and sorting said plurality of equine sperm cells based upon said sex.

79. The method of producing an equine animal as described in claim 78, further comprising the step of staining an amount of DNA contained within said plurality of equine sperm cells with a DNA selective material.

80. The method of producing an equine animal as described in claim 79, further comprising the steps of exposing said plurality of equine sperm cells containing said amount of DNA stained with said DNA selective material to a light source to generate an amount of fluorescence which varies based on the amount of said DNA stained with said DNA selective material.

81. The method of producing an equine animal as described in claim 80, further comprising the step of detecting said amount of fluorescence which varies based on the amount of said DNA stained with said DNA selective material.

82. The method of producing an equine animal as described in claim 81, wherein the step of staining an amount of DNA contained within said plurality of equine sperm cells with a DNA selective material occurs for a period of time sufficient to allow determination of said sex based on variation in said amount of fluorescence.

83. A method of producing an equine embryo of a selected sex as described in claim 69, wherein said step establishing discrete populations of equine sperm cells of same said sex provides a plurality of equine sperm cells having a purity selected from the group consisting of:

substantially one hundred percent, greater than ninety nine percent, greater than ninety five percent, greater than ninety percent, greater than eighty percent, greater than seventy percent, not less than seventy percent, not less than eighty percent, not less than ninety percent, not less than ninety five percent, not less than ninety nine percent, not less than one hundred percent, between about seventy percent and about eighty percent, between about eighty percent and about ninety percent, between about ninety percent and about one hundred percent.

84. An offspring foal having said selected sex produced by the method of claim 68.