US6146888A
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Method of enriching for mammalian stem cells
Assignee: University of Edinburgh
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Filing: 1994-04-21
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Granted: 2000-11-14
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Expires: 2014-04-21
Abstract
Abstract PCT No. PCT/GB94/00848 Sec. 371 Date Dec. 29, 1995 Sec. 102(e) Date Dec. 29, 1995 PCT Filed Apr. 21, 1994 PCT Pub. No. WO94/24274 PCT Pub. Date Oct. 27, 1994Mammalian stem cells are obtained and maintained in vitro whose genome has at least one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter specific for mammalian stem cells. The preferential expression of the antibiotic resistance gene in the stem cells results in the preferential survival of the stem cells in the presence of antibiotic.
Inventors
- 👤 Austin Gerard Smith
- 👤 Peter Scott Mountford
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 12:50
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/87; C12N15/90; C12N15/902; C12N15/907; A; A01; A01K; A01K67/00; A01K67/027; A01K67/0275; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4702; C07K14/52; C07K14/705; C07K14/70596; C07K14/81; C12N15/63; C12N15/79; C12N15/85; C12N15/8509; C12N15/873; C12N5/00; C12N5/06; C12N5/0602; C12N5/0618; C12N5/0623; C12N9/00; C12N9/10; C12N9/12; A01K2217/00; A01K2217/05; A01K2217/07; A01K2217/072; A01K2227/00; A01K2227/10; A01K2227/105; A01K2267/00; A01K2267/02; A61; A61K; A61K38/00; C12N2506/00; C12N2506/02; C12N2510/00; C12N2830/00; C12N2830/008; C12N2830/42; C12N2830/80; C12N2830/85; C12N2840/00; C12N2840/20; C12N2840/203; C12N2840/44
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
91 words
A method of enriching a population of mammalian cells for stem cells comprising: a) providing in vitro a mixed population of mammalian cells whose genome comprises at least one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter which preferentially expresses said antibiotic gene in mammalian stem cells, b) culturing said mixed population of mammalian cells in vitro under conditions conducive to cell survival wherein the preferential expression of said antibiotic resistance gene results in the preferential survival of mammalian stem cells in the presence of antibiotic.
Claim 2
Independent Claim
91 words
A method of enriching a population of mammalian cells for stem cells comprising: a) providing in vitro a mixed population of mammalian cells whose genome comprises at least one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter which preferentially expresses said antibiotic gene in mammalian stem cells, b) culturing said mixed population of mammalian cells in vitro under conditions conducive to cell survival wherein the preferential expression of said antibiotic resistance gene results in the preferential survival of mammalian stem cells in the presence of antibiotic.
Claim 3
Dependent Claim
25 words
The method according to claim 1 wherein the mammalian stem cells are selected from the group consisting of unipotential stem cells and pluripotential stem cells.
Claim 4
Dependent Claim
16 words
The method according to claim 1 wherein the antibiotic resistance gene is a neomycin resistance gene.
Claim 5
Dependent Claim
43 words
The method according to claim 1 wherein the genome of said mixed population of mammalian cells comprises two nucleic acid constructs each encoding an antibiotic resistance gene operatively linked to a promoter which preferentially expresses the antibiotic resistance gene in mammalian stem cells.
Claim 6
Dependent Claim
29 words
The method according to claim 1 wherein expression of the antibiotic resistance gene results from the introduction of a stably integrated, episomally maintained or transiently maintained nucleic acid construct.
Claim 7
Dependent Claim
32 words
The method according to claim 1 wherein expression of the antibiotic resistance gene results from operatively inserting the antibiotic resistance gene into an endogenous gene of the mixed population of mammalian cells.
Claim 8
Dependent Claim
31 words
The method according to claim 1 wherein the antibiotic resistance gene has been introduced into the mixed population of mammalian cells by transfection, lipofection, injection, ballistic missile, viral vector or electroporation.
Claim 9
Dependent Claim
25 words
The method according to claim 1 wherein the mixed population of cells is isolated from the group consisting of a cell line or an animal.
Claim 10
Dependent Claim
13 words
The method according to claim 1 wherein the promoter is an Oct4 promoter.
Claim 11
Dependent Claim
16 words
The method according to claim 1 wherein the antibiotic resistance gene is a neomycin phosphotransferase gene.
Claim 12
Dependent Claim
16 words
The method according to claim 1 wherein the promoter is preferentially expressed in hematopoietic stem cells.
Claim 13
Dependent Claim
13 words
The method according to claim 11 wherein the promoter is a CD34 promoter.
Claim 14
Dependent Claim
25 words
The method according to claim 1 wherein the mammalian stem cells are selected from the group consisting of embryonic stem cells and somatic stem cells.
Claim 15
Dependent Claim
25 words
The method according to claim 1 wherein the mammalian stem cells are selected from the group consisting of hematopoietic stem cells and neural stem cells.
Claim 16
Dependent Claim
32 words
An isolated mammalian stem cell comprising at one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter which preferentially expresses said antibiotic gene in said mammalian stem cell.
Claim 17
Dependent Claim
32 words
An isolated mammalian stem cell comprising at one nucleic acid construct encoding an antibiotic resistance gene operatively linked to a promoter which preferentially expresses said antibiotic gene in said mammalian stem cell.
References Cited (Backward Citations)
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