# US 2020/0399593 A1 · Method for obtaining a spermatozoid cell population with improved fitness > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399593 A1 - Document ID: US20200399593A1 - Status: Published application - Kind: A1 - Application number: 16/970,136 - Filed: 2019-02-14 - Published: 2020-12-24 - Earliest claimed: 2018-02-14 - Assignees: Universidad Del Pais Vasco/Euskal Herriko Unibertsitatea - Applicants: Universidad Del Pais Vasco/Euskal Herriko Unibertsitatea - Inventors: Nerea SUBIRÁN CIUDAD; Jon IRAZUSTA ASTIAZARAN; Luis CASIS SAENZ; Itziar URIZAR ARENAZA; Iraia MUÑOA HOYOS; Marta GIANZO CITORES - Web page: https://patentgrep.com/patent/US20200399593A1 ## CPC - C12N 5/0612: {sorting of gametes, e.g. according to sex or motility} - C12N 5/0604: {Whole embryos; Culture medium therefor} - C12N 5/061: {Sperm cells, spermatogonia} - G01N 33/56966: {Animal cells} - G01N 33/689: {related to pregnancy or the gonads} - C12N 2517/10: Conditioning of cells for in vitro fecondation or nuclear transfer - G01N 2333/70596: Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705 - G01N 2800/367: Infertility, e.g. sperm disorder, ovulatory dysfunction - 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 relates to a method for obtaining a spermatozoid cell population with improved fitness which comprises contacting the starting spermatozoid population with a binding agent recognizing the CD10 biomarker and isolating spermatozoids from the starting population which do not bind to said biomarker. The invention also relates to the spermatozoid cell population, which can be used to fertilize an ovum, and to the embryo obtained. The invention also relates to a method for determining the fitness of sperm for fertilization based on the percentage of the spermatozoid population not carrying the CD/10 biomarker. ## Claims ### Claim 1 A method for obtaining a spermatozoid cell population enriched in CD10− spermatozoids from a starting spermatozoid population comprising: (i) contacting the starting spermatozoid population with a CD10-binding agent and (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent. ### Claim 2 (depends on 1) The method according to claim 1 wherein the CD10-binding agent is an anti-CD10 antibody. ### Claim 3 (depends on 2) The method according to claim 2 wherein the isolation of CD10− spermatozoid is carried out by magnetic-activated cell sorting. ### Claim 4 (depends on 1, 2, 3) The method according to any of claims 1 to 3 wherein the starting spermatozoid population is isolated from a subject which is infertile, normozoospermic, asthenozoospermic, teratozoospermic, oligozoospermic or any combination thereof. ### Claim 5 (depends on 1, 2, 3, 4) The method according to any of claims 1 to 4 wherein the starting spermatozoid cell population is of a mammal. ### Claim 6 (depends on 5) The method according to claim 5 wherein the mammal is a human, porcine, bovine, equine, canine, murine or feline. ### Claim 7 A spermatozoid cell population enriched in CD10− spermatozoids. ### Claim 8 (depends on 7) The spermatozoid cell population according to claim 7 wherein the population derives from a mammal. ### Claim 9 (depends on 8) The spermatozoid cell population according to claim 8 wherein the mammal is a human, porcine, bovine, equine, canine, murine or feline. ### Claim 10 (depends on 7, 8, 9) An in vitro method for obtaining an embryo, said method selected from the group consisting of: (i) A method comprising fertilizing an ovum using a spermatozoid population according to any of claims 7 to 9 or (ii) A method comprising injecting into an ovum a spermatozoid from the spermatozoid cell population according to any of inventive claims 7 to 9 or (iii) A method comprising injecting into an ovum a CD10− spermatozoid. ### Claim 11 An in vitro method for obtaining an embryo comprising: (i) contacting a starting spermatozoid population with a CD10-binding agent, (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent thereby obtaining a spermatozoid cell population enriched in CD10− spermatozoids and (iii) fertilizing an ovum using a CD10− enriched spermatozoid cell population. ### Claim 12 An in vitro method for obtaining an embryo comprising: (i) contacting a starting spermatozoid population with a CD10-binding agent, (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent thereby obtaining a spermatozoid cell population enriched in CD10− spermatozoids and (iii) injecting into an ovum an spermatozoid from the CD10− enriched spermatozoid cell population obtained in step (ii). ### Claim 13 (depends on 11, 12) The method according to claim 11 or 12 wherein the CD10-binding agent is an anti-CD10 antibody. ### Claim 14 (depends on 13) The method according to claim 13 wherein the isolation of CD10− spermatozoid is carried out by magnetic-activated cell sorting. ### Claim 15 (depends on 11, 12, 13, 14) The method according to any of claims 11 to 14 wherein the starting spermatozoid population is isolated from a subject which is infertile, normozoospermic, asthenozoospermic, teratozoospermic, oligozoospermic or any combination thereof. ### Claim 16 (depends on 11, 12, 13, 14, 15) The method according to any of claims 11 to 15 wherein the starting spermatozoid cell population is of a mammal. ### Claim 17 (depends on 16) The method according to claim 16 wherein the mammal is a human, porcine, bovine, equine, canine, murine or feline. ### Claim 18 (depends on 7, 8, 9) An insemination method comprising administering into the reproductive tract of a female subject a spermatozoid population according to any of claims 7 to 9. ### Claim 19 An insemination method comprising: (i) contacting a starting spermatozoid population with a CD10-binding agent, (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent thereby obtaining a spermatozoid cell population enriched in CD10− spermatozoids and (iii) administering into the reproductive tract of a female subject the spermatozoid cell population obtained in step (ii). ### Claim 20 (depends on 19) The method according to claim 19 wherein the CD10-binding agent is an anti-CD10 antibody. ### Claim 21 (depends on 20) The method according to claim 20 wherein the isolation of CD10− spermatozoid is carried out by magnetic-activated cell sorting. ### Claim 22 (depends on 19, 20, 21) The method according to any of claims 19 to 21 wherein the starting spermatozoid population is isolated from a subject which is infertile, normozoospermic, asthenozoospermic, teratozoospermic, oligozoospermic or any combination thereof. ### Claim 23 (depends on 19, 20, 21, 22) The method according to any of claims 19 to 22 wherein the starting spermatozoid cell population is of a mammal. ### Claim 24 (depends on 23) The method according to claim 23 wherein the mammal is a human, porcine, bovine, equine, canine, murine or feline. ### Claim 25 (depends on 19, 20, 21, 22, 23, 24) The insemination method according to any of claims 19 to 24 wherein the spermatozoid population is administered in the uterus of the female subject. ### Claim 26 (depends on 8, 9, 10, 11, 12, 13, 14, 15, 16, 17) An embryo obtained by in vitro fertilization of an ovum with a spermatozoid population according to any of claims 8 to 10 or by a method as defined in any of claims 10 to 17. ### Claim 27 (depends on 26) The embryo according to claim 26 wherein the fertilization is achieved by ICSI (Intracytoplasmic Sperm Injection). ### Claim 28 (depends on 26, 27) The embryo according to any one of claim 26 or 27 which is non-human. ### Claim 29 (depends on 26, 27, 28) The embryo according to claims 26 to 28 wherein the embryo is in the blastocyst stage. ### Claim 30 (depends on 26, 27, 28, 29) A method for obtaining a blastocyst with an increased implantation rate comprising obtaining or providing an embryo as defined in any one of claims 26 to 29 and allowing the embryo to develop until the stage of blastocyst. ### Claim 31 A method for obtaining a blastocyst with an increased implantation rate comprising (i) contacting a starting spermatozoid population with a CD10-binding agent, (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent thereby obtaining a spermatozoid cell population enriched in CD10− spermatozoids, (iii) fertilizing an ovum using a CD10− enriched spermatozoid cell population and (iv) allowing the embryo to develop until the stage of blastocyst. ### Claim 32 A method for obtaining a blastocyst with an increased implantation rate comprising: (i) contacting a starting spermatozoid population with a CD10-binding agent, (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent thereby obtaining a spermatozoid cell population enriched in CD10− spermatozoids and (iii) injecting into an ovum an spermatozoid from the CD10− enriched spermatozoid cell population obtained in step (ii) and (iv) allowing the embryo to develop until the stage of blastocyst. ### Claim 33 (depends on 26, 27, 28, 29) A method for achieving pregnancy in a female subject comprising transferring into said female subject an embryo according to any of claims 26 to 29. ### Claim 34 A method for achieving pregnancy in a female subject comprising: (i) contacting a starting spermatozoid population with a CD10-binding agent, (ii) isolating spermatozoids from the starting population which do not bind to the CD10-binding agent thereby obtaining a spermatozoid cell population enriched in CD10− spermatozoids, (iii) fertilizing an ovum using the CD10− enriched spermatozoid cell population obtained in step (iii) and (iv) transferring into said female subject the embryo obtained in step (iii). ### Claim 35 (depends on 32) The method according to claim 32 wherein the CD10-binding agent is an anti-CD10 antibody. ### Claim 36 (depends on 34) The method according to claim 34 wherein the isolation of CD10− spermatozoid is carried out by magnetic-activated cell sorting. ### Claim 37 (depends on 34, 35, 36) The method according to any of claims 34 to 36 wherein the starting spermatozoid population is isolated from a subject which is infertile, normozoospermic, asthenozoospermic, teratozoospermic, oligozoospermic or any combination thereof. ### Claim 38 (depends on 34, 35, 36, 37) The method according to any of claims 34 to 37 wherein the starting spermatozoid cell population is of a mammal. ### Claim 39 (depends on 38) The method according to claim 38 wherein the mammal is a human, porcine, bovine, equine, canine, murine or feline. ### Claim 40 (depends on 34, 35, 36, 37, 38, 39) A method for achieving pregnancy in a female subject according to any of claims 34 to 39 wherein said female is infertile. ### Claim 41 (depends on 34, 35, 36, 37, 38, 39, 40) The method according to claims 34 to 40 wherein the embryo that is to be transferred into the female mammal is a blastocyst. ### Claim 42 Use of a CD10-binding agent for the preparation of a spermatozoid cell population enriched in CD10− spermatozoids. ### Claim 43 Use of a CD10-binding agent for the preparation of a spermatozoid cell population having a high degree of DNA integrity. ### Claim 44 Use of a CD10-binding agent for the preparation of a spermatozoid cell population having a low level of aneuploidies. ### Claim 45 (depends on 42, 43, 44) The use according to any of claims 42 to 44 wherein the CD10-binding agent is an anti-CD10 antibody. ### Claim 46 A method for determining the fitness of, the DNA integrity of or the number of aneuplodies in a human sperm population for fertilization which comprises determining the content of CD10− spermatozoids in a said population, wherein increased levels of CD10− spermatozoids in the population with respect to a reference value are indicative that the human sperm is adequate for fertilization, that the sperm population shows a high degree of DNA integrity or a low level of aneuploidies. ### Claim 47 (depends on 46) The method according to claim 46, wherein the content of CD10− spermatozoids is determined with a CD10-binding agent. ### Claim 48 (depends on 47) The method according to claim 47 wherein the CD10-binding agent is an antibody. ### Claim 49 (depends on 46, 47, 48) The method according to any of claims 46 to 48 wherein the human sperm is isolated from a subject which is infertile, normozoospermic, asthenozoospermic, teratozoospermic, oligozoospermic or any combination thereof. ### Claim 50 (depends on 46, 47, 48, 49) The method according to any of claims 46 to 49 wherein the human sperm population is a semen sample.