US6235970B1
Expired / Lapsed
US
⏱️ 0.0 years remaining
CICM cells and non-human mammalian embryos prepared by nuclear transfer of a proliferating differentiated cell or its nucleus
Assignee: University of Massachusetts Amherst
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Filing: 1997-09-22
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Granted: 2001-05-22
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Expires: 2017-09-22
Abstract
Abstract An improved method of nuclear transfer involving the transplantation of donor differentiated cell nuclei into enucleated oocytes of the same species as the donor cell is provided. The resultant nuclear transfer units are useful for multiplication of genotypes and transgenic genotypes by the production of fetuses and offspring, and for production of isogenic CICM cells, including human isogenic embryonic or stem cells. Production of genetically engineered or transgenic mammalian embryos, fetuses and offspring is facilitated by the present method since the differentiated cell source of the donor nuclei can be genetically modified and clonally propagated.
Inventors
- 👤 Steven L. Stice
- 👤 Jose Cibelli
- 👤 James Robl
- 👤 Paul Golueke
- 👤 F. Abel Ponce de Leon
- 👤 D. Joseph Jerry
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 05:02
CPC Classifications
A; A01; A01K; A01K67/00; A01K67/027; A01K67/0275; A61; A61P; A61P1/00; A61P1/18; A61P13/00; A61P13/02; A61P17/00; A61P17/02; A61P21/00; A61P25/00; A61P25/14; A61P25/16; A61P25/28; A61P3/00; A61P3/08; A61P3/10; A61P31/00; A61P31/12; A61P31/14; A61P31/18; A61P35/00; A61P9/00; A61P9/04; C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; C12N15/873; C12N15/877; C12N15/8771; C12N15/8778; C12N5/00; C12N5/06; C12N5/0602; C12N5/0603; A01K2217/00; A01K2217/05; A01K2227/00; A01K2227/10; A01K2227/101; A01K2227/106; A61K; A61K35/00; A61K35/12; C12N2510/00; C12N2517/00; C12N2517/02; C12N2517/04
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
102 words
A method for producing a mammalian cultured inner cell mass (CICM) cell line by nuclear transfer comprising the following steps: (i) introducing a proliferating differentiated somatic mammalian donor cell or a proliferating differentiated somatic mammalian donor cell nucleus into an enucleated mammalian oocyte of the same species to produce a nuclear transfer unit; (ii) activating the resultant nuclear transfer unit; (iii) culturing said activated nuclear transfer unit until at least a size suitable for obtaining ICM cells: (iv) isolating and culturing ICM cells obtained from said cultured nuclear transfer unit to obtain a cultured inner cell mass (CICM) or CICM cell line.
Claim 2
Independent Claim
102 words
A method for producing a mammalian cultured inner cell mass (CICM) cell line by nuclear transfer comprising the following steps: (i) introducing a proliferating differentiated somatic mammalian donor cell or a proliferating differentiated somatic mammalian donor cell nucleus into an enucleated mammalian oocyte of the same species to produce a nuclear transfer unit; (ii) activating the resultant nuclear transfer unit; (iii) culturing said activated nuclear transfer unit until at least a size suitable for obtaining ICM cells: (iv) isolating and culturing ICM cells obtained from said cultured nuclear transfer unit to obtain a cultured inner cell mass (CICM) or CICM cell line.
Claim 3
Dependent Claim
32 words
The method of claim 1 , wherein said culturing step (iii) is effected on a feeder layer, and results in the production of a NT unit having at least about 50 cells.
Claim 4
Dependent Claim
65 words
The method of claim 2 , wherein said donor cell or donor cell nucleus is selected from the group consisting of epithelial cells, neural cells, epidermal cells, keratinocytes, hematopoietic cells, melanocytes, chondrocytes, lymphocytes, erythrocytes, macrophages, monocytes, mononuclear cells, fibroblasts, muscle cells, skin cells, lung cells, pancreatic cells, liver cells, stomach cells, intestinal cells, heart cells, bladder cells, reproductive organ cells, urethra cell, and kidney cells.
Claim 5
Dependent Claim
36 words
The method of claim 1 , wherein step (iv) comprises culturing said NT unit to form a blastocyst, and culturing ICM cells obtained from said blastocyst to produce cultured inner cell mass (CICM) or cell lines.
Claim 6
Dependent Claim
17 words
The method of claim 1 , wherein said donor cell or donor cell nucleus is genetically modified.
Claim 7
Dependent Claim
23 words
The method of claim 5 , wherein the genome of said genetically modified cell comprises at least one insertion, deletion or substitution modification.
Claim 8
Dependent Claim
20 words
The method of claim 6 , wherein said genetically modified donor cell or donor cell nucleus comprises a heterologous DNA.
Claim 9
Dependent Claim
42 words
The method of claim 6 , wherein at least one said gene or a portion thereof in said genetically modified donor cell or donor cell nucleus has been inserted, deleted or modified at a specific site in the genome of said cell.
Claim 10
Dependent Claim
21 words
The method of claim 8 , wherein said genetically modified donor cell or donor cell nucleus is produced by homologous recombination.
Claim 11
Dependent Claim
36 words
The method of claim 1 , wherein said donor cell or donor cell nucleus is isolated from a mammal selected from the group consisting of sheep, cows, pigs, horses, rabbits, guinea pigs, hamsters, rats and primates.
Claim 12
Dependent Claim
20 words
The method of claim 10 , wherein said donor cell or donor cell nucleus comprises at least one genetic modification.
Claim 13
Dependent Claim
18 words
The method of claim 10 , wherein said donor cell or donor cell nucleus is a human cell.
Claim 14
Dependent Claim
19 words
The method of claim 1 , wherein said donor cell or donor cell neucleus is isolated from an ungulate.
Claim 15
Dependent Claim
18 words
The method of claim 1 , wherein said donor cell or donor cell nucleus is a human cell.
Claim 16
Dependent Claim
18 words
The method of claim 1 , wherein said donor cell is expanded in vitro prior to step (i).
Claim 17
Dependent Claim
38 words
The method of claim 1 , wherein in step (iv) the ICM cells of said cultured NT unit are transferred onto a feeder layer and cultured to produce a cultured inner cell mass (CICM) or CICM cell line.
Claim 18
Dependent Claim
15 words
The method of claim 16 , wherein the feeder layer is a fibroblast feeder layer.
Claim 19
Dependent Claim
21 words
The method of claim 1 , wherein the cultured inner cell mass (CICM) or CICM cell line is bovine or porcine.
Claim 20
Independent Claim
87 words
A method of producing a non-human mammalian embryo by nuclear transfer comprising transplantation of a non-human mammalian cell or a nucleus of a non-human mammalian cell into an enucleated oocyte of the same species as the donor cell or donor cell nucleus, activation of the recipient oocyte containing the donor cell or donor cell nucleus, and incubation of the activated oocyte to produce an embryo, wherein the donor cell is a proliferating mammalian differentiated cell or wherein the donor nucleus is from a proliferating mammalian differentiated cell.
Claim 21
Independent Claim
87 words
A method of producing a non-human mammalian embryo by nuclear transfer comprising transplantation of a non-human mammalian cell or a nucleus of a non-human mammalian cell into an enucleated oocyte of the same species as the donor cell or donor cell nucleus, activation of the recipient oocyte containing the donor cell or donor cell nucleus, and incubation of the activated oocyte to produce an embryo, wherein the donor cell is a proliferating mammalian differentiated cell or wherein the donor nucleus is from a proliferating mammalian differentiated cell.
Claim 22
Dependent Claim
30 words
The method according to claim 19 wherein said non-human mammalian embryo is selected from the group consisting of sheep, cows, pigs, horses, rabbits, guinea pigs, hamsters, rats and non-human primates.
Claim 23
Dependent Claim
14 words
The method according to claim 19 wherein said non-human mammalian embryo is an ungulate.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2004-05-21 | 2004-11-20 | 2005-05-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-05-21 | 2008-11-20 | 2009-05-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-05-21 | 2012-11-20 | 2013-05-22 | $7700.00 | Window Closed / Lapsed |