Strategy for maintaining pregnancy

• US 6,673,987 B1
• Filed: 03/29/2000
• Issued: 01/06/2004
• Est. Priority Date: 03/24/2000
• Status: Expired due to Fees

• Current Assignee: Geron Corporation
• Original Assignee: Geron Corporation
• Inventors: King, Timothy J.
• Primary Examiner(s): Crouch, Deborah
• Assistant Examiner(s): WOITACH, JOSEPH T
• Application Number:
  US09/538,867
• Time in Patent Office: 1,378 Days
• Field of Search: 800/14, 800/3, 800/4, 800/8, 800/21
• US Class Current: 800/21
• CPC Class Codes: A61K 2035/122 for inducing tolerance or supression of immune responses
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61P 15/06 Antiabortive agents; Labour repressants
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• CPC Class Codes: C12N 5/0604 Whole embryos; Culture medium therefor

Abstract

Pregnancy in animals such as the pig usually does not continue to term when there are only a few embryos in the uterus. Currently available technology for maintaining pregnancy involves a series of hormone injections, and the failure rate is high. It has now been discovered that pregnancy can be maintained by complementing fertile embryos in the uterus with additional embryos that aren'"'"'t expected to survive the full term of the pregnancy. Particularly suitable are parthenogenetic embryos, formed by activating and diploidizing unfertilized oocytes. The carrier embryos are engrafted into the uterus of a surrogate female, and the pregnancy continues to term without further intervention—even if the number of fertile embryos are below the minimum litter size. This provides valuable biological material that can be used for transplantation, the production of pharmaceuticals, and for agricultural use.

Claims

1. A method for producing a non-human mammal in a species wherein maintenance of pregnancy depends on having a sufficient plurality of embryos in the uterus, comprising:

a) combining at least one embryo produced by nuclear transfer with a plurality of infertile embryos of the same species produced by a method that does not involve nuclear transfer;

b) engrafting the combined embryos into the uterus of a pseudo-pregnant female of the same species; and

subsequently

c) birthing a mammal from the female resulting from intrauterine development of the embryo produced by nuclear transfer.

2. The method of claim 1, wherein the infertile embryos are parthenogenetic embryos.

3. The method of claim 2, wherein the parthenogenetic embryos are produced by diploidizing and activating an oocyte of the same species as the female.

4. The method of claim 1, wherein the infertile embryos comprise cells with abnormal ploidy.

5. The method of claim 1, wherein at least five infertile embryos are engrafted into the uterus.

6. The method of claim 1, wherein the embryo produced by nuclear transfer contains chromosomal DNA that has been genetically altered.

7. The method of claim 6, wherein the chromosomal DNA has been genetically altered to express a heterologous gene.

8. The method of claim 6, wherein the chromosomal DNA has been genetically altered to prevent expression of a gene that would otherwise be expressed.

9. The method of claim 1, wherein the species is a domestic swine.

10. A method for producing a non-human mammal in a species wherein maintenance of pregnancy depends on having a sufficient plurality of embryos in the uterus, comprising:

a) engrafting at least one embryo produced by nuclear transfer into the uterus of a female of the same species;

b) engrafting into the uterus of the female a plurality of infertile embryos of the same species produced by a method that does not involve nuclear transfer at a time before or after a) so as to prevent regression of the corpora lutea and loss of the embryo produced by nuclear transfer; and

subsequently

c) birthing a mammal from the female resulting from intrauterine development of the embryo produced by nuclear transfer.

11. The method of claim 10, Wherein the infertile embryos are parthenogenetic embryos.

12. The method of claim 11, wherein the parthenogenetic embryos are produced by diploidizing and activating an oocyte of the same species as the female.

13. The method of claim 10, wherein the infertile embryos comprise cells with abnormal ploidy.

14. The method of claim 10, wherein at least five infertile embryos are engrafted into the uterus.

15. The method of claim 10, wherein the embryo produced by nuclear transfer contains DNA that has been genetically altered.

16. The method of claim 15, wherein the chromosomal DNA has been genetically altered to express a heterologous gene.

17. The method of claim 15, wherein the chromosomal DNA has been genetically altered to prevent expression of a gene that would otherwise be expressed.

18. The method of claim 10, wherein the species is a domestic swine.

19. A method for maintaining pregnancy in a pig, comprising engrafting the pig with at least one fertile embryo plus one or more parthenogenetic embryos, whereby the number of fertile embryo(s) and parthenogenetic embryo(s) is sufficient to prevent regression of the corpora lutea, thereby allowing pregnancy to be maintained such that a mammal is produced from development of the fertile embryo in utero.

20. The method for maintaining pregnancy of claim 19, wherein the fertile embryo contains chromosomal DNA that has been genetically altered to include an expressible encoding region for a human protein, or to inactivate a gene that would otherwise be expressed.

21. A pig carrying at least one fertile embryo, plus one or more artificially engrafted embryos produced by parthenogenetic activation of an oocyte without nuclear transfer.

22. The pig of claim 21, wherein the fertile embryo contains chromosomal DNA that has been genetically altered to include an expressible encoding region for a human protein, or to inactivate a gene that would otherwise be expressed.