US5639618A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method of isolating a lineage specific stem cell in vitro
Assignee: Plurion Inc
•
Filing: 1994-05-13
•
Granted: 1997-06-17
•
Expires: 2014-05-13
Abstract
Abstract The present invention provides a method of isolating a lineage specific stem cell in vitro, comprising a) transfecting a pluripotent embryonic stem cell with a construct comprising a regulatory region of a lineage specific gene operably linked to a DNA encoding a reporter protein, b) culturing the pluripotent embryonic stem cell under conditions such that the pluripotent embryonic stem cell differentiates into a lineage specific stem cell and c) separating the cells which express the reporter protein from the other cells in the culture, the cell which expresses the reporter protein being an isolated lineage specific stem cell. A lineage specific stem cell can also be identified utilizing this method.
Inventors
- 👤 David A. Gay
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 22:43
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0618; C12N5/0623; C12N15/00; C12N15/09; C12N15/63; C12N15/65; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/569; G01N33/56966; C12N2501/00; C12N2501/60; C12N2506/00; C12N2506/02; C12N2510/00
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
105 words
A method of isolating a lineage specific stem cell in vitro, comprising: a. transfecting in vitro a pluripotent embryonic stem cell with a construct comprising a regulatory region of a lineage specific gene operably linked to a DNA encoding a reporter protein; b. culturing the pluripotent embryonic stem cell in vitro under conditions such that the pluripotent embryonic stem cell differentiates into a lineage specific stem cell; and c. separating the cells fluorescent activated cell sorting or by immunoaffinity procedures which express the reporter protein from the other cells in culture, the cell which expresses the reporter protein being an isolated lineage specific stem cell.
Claim 2
Independent Claim
105 words
A method of isolating a lineage specific stem cell in vitro, comprising: a. transfecting in vitro a pluripotent embryonic stem cell with a construct comprising a regulatory region of a lineage specific gene operably linked to a DNA encoding a reporter protein; b. culturing the pluripotent embryonic stem cell in vitro under conditions such that the pluripotent embryonic stem cell differentiates into a lineage specific stem cell; and c. separating the cells fluorescent activated cell sorting or by immunoaffinity procedures which express the reporter protein from the other cells in culture, the cell which expresses the reporter protein being an isolated lineage specific stem cell.
Claim 3
Dependent Claim
31 words
The method of claim 1 wherein the regulatory region of the lineage specific gene is selected from the group consisting of an Otx, Dlx, NIx, Emx, Wnt, En, Hox and ACHRβ.
Claim 4
Dependent Claim
17 words
The method of claim 1 wherein the reporter protein is a protein expressed on the cell surface.
Claim 5
Dependent Claim
11 words
The method of claim 1 wherein the reporter protein is LacZ.
Claim 6
Independent Claim
102 words
A method of isolating a lineage specific stem cell in vitro, comprising: a. transfecting in vitro a pluripotent embryonic stem cell with a construct comprising a regulatory region of a lineage specific gene operably linked to a DNA encoding a reporter protein; b. culturing the pluripotent embryonic stem cell in vitro under conditions which allow the formation of aggregates of differentiating cells; and c. separating the cells by fluorescent activated cell sorting or by immunoaffinity procedures in the aggregates which express the reporter protein from the other cells, the cell which expresses the reporter protein being an isolated lineage specific stem cell.
Claim 7
Independent Claim
102 words
A method of isolating a lineage specific stem cell in vitro, comprising: a. transfecting in vitro a pluripotent embryonic stem cell with a construct comprising a regulatory region of a lineage specific gene operably linked to a DNA encoding a reporter protein; b. culturing the pluripotent embryonic stem cell in vitro under conditions which allow the formation of aggregates of differentiating cells; and c. separating the cells by fluorescent activated cell sorting or by immunoaffinity procedures in the aggregates which express the reporter protein from the other cells, the cell which expresses the reporter protein being an isolated lineage specific stem cell.
Claim 8
Dependent Claim
19 words
The method of claim 5 wherein the aggregates are cultured to form embryoid bodies prior to separating the cells.
Claim 9
Dependent Claim
31 words
The method of claim 5 wherein the regulatory region of the lineage specific gene is selected from the group consisting of an Otx, Dlx, Nlx, Emx, Wnt, En, Hox and ACHRβ.
Claim 10
Dependent Claim
15 words
The method of claim 5 wherein the reporter protein is expressed on the cell surface.
Claim 11
Dependent Claim
11 words
The method of claim 5 wherein the reporter protein is LacZ.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2000-06-16 | 2000-12-16 | 2001-06-17 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2004-06-16 | 2004-12-16 | 2005-06-17 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2008-06-16 | 2008-12-16 | 2009-06-17 | $7700.00 | Window Closed / Lapsed |