# US 8,771,934 B2 · Inorganic pyrophosphate and uses thereof

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- Number: US 8,771,934 B2
- Document ID: US8771934B2
- Status: Granted
- Kind: B2
- Application number: 13/708,728
- Filed: 2012-12-07
- Published: 2013-06-27
- Granted: 2014-07-08
- Earliest claimed: 2011-12-09
- Assignees: The Curators of the University of Missouri
- Applicants: The Curators of the University of Missouri
- Inventors: Peter Sutovsky; Young-Joo Yi
- Web page: https://patentgrep.com/patent/US8771934B2

## CPC

- A61B 17/43: for artificial insemination
- A01N 1/126: Physiologically active agents, e.g. antioxidants or nutrients
- A61D 19/02: for artificial insemination
- C12N 5/0604: {Whole embryos; Culture medium therefor}
- C12N 5/0609: {Oocytes, oogonia (fertilised oocytes C12N5/0604)}
- C12N 5/061: {Sperm cells, spermatogonia}
- C12N 5/0612: {sorting of gametes, e.g. according to sex or motility}
- G01N 33/5005: {involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025)}
- C12N 2500/05: Inorganic components
- C12N 2517/10: Conditioning of cells for in vitro fecondation or nuclear transfer
- A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D)
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A61D: VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The present invention provides a new and improved sperm stimulating additive comprising a certain amount of inorganic pyrophosphate (PPi). Addition of PPi in the media for human/animal in vitro fertilization (IVF) improves fertilization rate; addition of PPi in the semen extender for farm animal artificial insemination (AI) may improve pregnancy rates; furthermore, mammalian oocytes matured in vitro in a medium including PPi attain improved fertilization and developmental potential, while embryos cultured in medium supplemented with PPi have improved development to blastocyst.

## Claims

### Claim 1

A sperm preservation media comprising inorganic pyrophosphate (PPi) and at least one spermatozoa, wherein the concentration of PPi is between about 1 μM and about 200 μM.

### Claim 2 (depends on 1)

The sperm preservation media of claim 1, wherein the concentration of PPi is between about 1 μM and about 20 μM.

### Claim 3 (depends on 1)

The sperm preservation media of claim 1, wherein the concentration of PPi is about 10 μM.

### Claim 4 (depends on 1)

The sperm preservation media of claim 1, wherein said preservation media is used to preserve sperm from a porcine.

### Claim 5

A media for sperm processing comprising inorganic pyrophosphate (PPi) and at least one spermatozoa, wherein the concentration of PPi is between about 1 μM and about 200 μM.

### Claim 6 (depends on 5)

The media of claim 5, wherein the concentration of PPi is between about 1 μM and about 20 μM.

### Claim 7

A media for in vitro fertilization (IVF) or artificial insemination (AI) comprising inorganic pyrophosphate (PPi) and at least one spermatozoa, wherein the concentration of PPi is between about 1 μM and about 200 μM.

### Claim 8 (depends on 7)

The media of claim 7, wherein the concentration of PPi is between about 1 μM and about 20 μM.

### Claim 9

A method of sperm preservation comprising storing sperm in a media comprising inorganic pyrophosphate (PPi), wherein the concentration of PPi is between about 1 μM to about 200 μM.

### Claim 10 (depends on 9)

The method of claim 9, wherein the concentration of PPi is between about 1 μM to about 20 μM.

### Claim 11 (depends on 9)

The method of claim 9, wherein the concentration of PPi is about 10 μM.

### Claim 12 (depends on 9)

The method of claim 9, wherein the sperm is stored in the media comprising PPi for up to 10 days.