US5843780A
Expired / Lapsed
US
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Primate embryonic stem cells
Assignee: Wisconsin Alumni Research Foundation
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Filing: 1996-01-18
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Granted: 1998-12-01
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Expires: 2016-01-18
Abstract
Abstract A purified preparation of primate embryonic stem cells is disclosed. This preparation is characterized by the following cell surface markers: SSEA-1 (-); SSEA-3 (+); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); and alkaline phosphatase (+). In a particularly advantageous embodiment, the cells of the preparation have normal karyotypes and continue to proliferate in an undifferentiated state after continuous culture for eleven months. The embryonic stem cell lines also retain the ability, throughout the culture, to form trophoblast and to differentiate into all tissues derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). A method for isolating a primate embryonic stem cell line is also disclosed.
Inventors
- 👤 James A. Thomson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 14:59
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0603; C12N5/0605; A; A61; A61P; A61P25/00; A61P25/14; A61P25/16; A61P3/00; A61P3/08; A61P3/10; A61P37/00; C12N5/0606; C12N2502/00; C12N2502/13; C12N2506/00; C12N2506/02
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
75 words
A purified preparation of primate embryonic stem cells which (i) is capable of proliferation in an in vitro culture for over one year, (ii) maintains a karyotype in which all the chromosomes characteristic of the primate species are present and not noticeably altered through prolonged culture, (iii) maintains the potential to differentiate into derivatives of endoderm, mesoderm, and ectoderm tissues throughout the culture, and (iv) will not differentiate when cultured on a fibroblast feeder layer.
Claim 2
Dependent Claim
75 words
A purified preparation of primate embryonic stem cells which (i) is capable of proliferation in an in vitro culture for over one year, (ii) maintains a karyotype in which all the chromosomes characteristic of the primate species are present and not noticeably altered through prolonged culture, (iii) maintains the potential to differentiate into derivatives of endoderm, mesoderm, and ectoderm tissues throughout the culture, and (iv) will not differentiate when cultured on a fibroblast feeder layer.
Claim 3
Dependent Claim
23 words
The preparation of claim 1 wherein the stem cells will spontaneously differentiate to trophoblast and produce chorionic gonadotropin when cultured to high density.
Claim 4
Dependent Claim
60 words
A purified preparation of primate embryonic stem cells wherein the cells are negative for the SSEA-1 marker, positive for the SSEA-3 marker, positive for the SSEA-4 marker, express alkaline phosphatase activity, are pluripotent, and have karyotypes which includes the presence of all of the chromosomes characteristic of the primate species and in which none of the chromosomes are noticeably altered.
Claim 5
Dependent Claim
60 words
A purified preparation of primate embryonic stem cells wherein the cells are negative for the SSEA-1 marker, positive for the SSEA-3 marker, positive for the SSEA-4 marker, express alkaline phosphatase activity, are pluripotent, and have karyotypes which includes the presence of all of the chromosomes characteristic of the primate species and in which none of the chromosomes are noticeably altered.
Claim 6
Dependent Claim
16 words
The preparation of claim 3 wherein the cells are positive for the TRA-1-60, and TRA-1-81 markers.
Claim 7
Dependent Claim
23 words
The preparation of claim 3 wherein the cells continue to proliferate in an undifferentiated state after continuous culture for at least one year.
Claim 8
Dependent Claim
21 words
The preparation of claim 3 wherein the cells will differentiate to trophoblast when cultured beyond confluence and will produce chorionic gonadotropin.
Claim 9
Dependent Claim
18 words
The preparation of claim 3 wherein the cells remain euploid for more than one year of continuous culture.
Claim 10
Dependent Claim
28 words
The preparation of claim 3 wherein the cells differentiate into cells derived from mesoderm, endoderm and ectoderm germ layers when the cells are injected into a SCID mouse.
Claim 11
Independent Claim
92 words
A method of isolating a primate embryonic stem cell line, comprising the steps of: (a) isolating a primate blastocyst; (b) isolating cells from the inner cell mass of the blastocyst of (a); (c) plating the inner cell mass cells on embryonic fibroblasts, wherein inner cell mass-derived cells masses are formed; (d) dissociating the mass into dissociated cells; (e) replating the dissociated cells on embryonic feeder cells; (f) selecting colonies with compact morphologies and cells with high nucleus to cytoplasm ratios and prominent nucleoli; and (g) culturing the cells of the selected colonies.
Claim 12
Independent Claim
92 words
A method of isolating a primate embryonic stem cell line, comprising the steps of: (a) isolating a primate blastocyst; (b) isolating cells from the inner cell mass of the blastocyst of (a); (c) plating the inner cell mass cells on embryonic fibroblasts, wherein inner cell mass-derived cells masses are formed; (d) dissociating the mass into dissociated cells; (e) replating the dissociated cells on embryonic feeder cells; (f) selecting colonies with compact morphologies and cells with high nucleus to cytoplasm ratios and prominent nucleoli; and (g) culturing the cells of the selected colonies.
Claim 13
Dependent Claim
21 words
A method as claimed in claim 9 further comprising maintaining the isolated cells on a fibroblast feeder layer to prevent differentiation.
Claim 14
Dependent Claim
10 words
A cell line developed by the method of step 9.
Claim 15
Dependent Claim
10 words
A cell line developed by the method of step 9.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-11-30 | 2002-06-01 | 2002-12-01 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-11-30 | 2006-06-01 | 2006-12-01 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-11-30 | 2010-06-01 | 2010-12-01 | $7700.00 | Window Closed / Lapsed |