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US5166065A Expired / Lapsed US ⏱️ 0.0 years remaining

In vitro propagation of embryonic stem cells

Assignee: CSL Innovation Pty Ltd
•
Filing: 1989-08-03
•
Granted: 1992-11-24
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Expires: 2009-08-03
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Abstract

Abstract PCT No. PCT/AU89/00330 Sec. 371 Date May 31, 1990 Sec. 102(e) Date May 31, 1990 PCT Filed Aug. 3, 1989 PCT Pub. No. WO90/01541 PCT Pub. Date Feb. 22, 1990.The present invention relates generally to the use of leukaemia inhibitory factor (LIF) in the maintenance and derivation of embryonic stem (ES) cells in culture. The ES cells are maintained and/or derived from animal embryos by culturing said cells or embryos in a culture medium containing an effective amount of LIF for a time and under conditions sufficient to maintain and/or derive said ES cells. The ES cells may be passaged in LIF and used to make chimaeric animals.

Inventors

  • 👤 Robert L. Williams
  • 👤 Nicholas M. Gough
  • 👤 Douglas J. Hilton

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-03 10:56
CPC Classifications
A; A01; A01K; A01K67/00; A01K67/027; A01K67/0275; C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0603; C12N5/0606; A01K2217/00; A01K2217/05; C12N2501/00; C12N2501/20; C12N2501/23; C12N2501/235; C12N2510/00

Claims Analysis

Total Claims: 18
Claim 1
Independent Claim 50 words
A method for the isolation of embryonic stem (ES) cells from mammalian embryos in vitro which method comprises deriving and maintaining said embryos in culture medium containing an effective amount of recombinant leukaemia inhibitory factor (LIF) for a time and under conditions sufficient for the development of said ES cells.
Claim 2
Independent Claim 50 words
A method for the isolation of embryonic stem (ES) cells from mammalian embryos in vitro which method comprises deriving and maintaining said embryos in culture medium containing an effective amount of recombinant leukaemia inhibitory factor (LIF) for a time and under conditions sufficient for the development of said ES cells.
Claim 3
Dependent Claim 15 words
The method according to claim 1 wherein the culture medium is free of feeder cells.
Claim 4
Dependent Claim 19 words
The method according to claim 1 wherein the culture medium is Eagle's medium or modifications thereof or equivalent thereto.
Claim 5
Dependent Claim 15 words
The method according to claim 1 wherein the LIF is recombinant human or murine LIF.
Claim 6
Dependent Claim 23 words
The method according to claim 4 wherein LIF is added to the culture medium at a concentration of from 10 to 1,000,000 units/ml.
Claim 7
Dependent Claim 24 words
The method according to claim 5 wherein the LIF is added to the culture medium at a concentration of from 100 to 100,000 units/ml.
Claim 8
Dependent Claim 24 words
The method according to claim 6 wherein the LIF is added to the culture medium at a concentration of from 500 to 10,000 units/ml.
Claim 9
Dependent Claim 17 words
A method according to claim 1 wherein the effective time is from 1 day to 20 weeks.
Claim 10
Dependent Claim 16 words
The method according to claim 8 wherein the effective time is from 1 to 8 weeks.
Claim 11
Independent Claim 43 words
A method for maintaining mammalian embryonic stem (ES) cells in vitro while retaining their pluripotential phenotype which process comprises culturing said cells in a culture medium containing an effective amount of recombinant leukaemia inhibitory factor (LIF) under conditions sufficient to maintain said cells.
Claim 12
Independent Claim 43 words
A method for maintaining mammalian embryonic stem (ES) cells in vitro while retaining their pluripotential phenotype which process comprises culturing said cells in a culture medium containing an effective amount of recombinant leukaemia inhibitory factor (LIF) under conditions sufficient to maintain said cells.
Claim 13
Dependent Claim 15 words
The method according to claim 10 wherein the culture medium is free of feeder cells.
Claim 14
Dependent Claim 19 words
The method according to claim 10 wherein the culture medium comprises Eagle's medium or modifications thereof or equivalents thereto.
Claim 15
Dependent Claim 15 words
The method according to claim 10 wherein the LIF is recombinant murine or human LIF.
Claim 16
Dependent Claim 25 words
The method according to claim 13 wherein the recombinant LIF is added to the culture medium at a concentration of from 10 to 1,000,000 units/ml.
Claim 17
Dependent Claim 25 words
The method according to claim 14 wherein the recombinant LIF is added to the culture medium at a concentration of from 100 to 100,000 units/ml.
Claim 18
Dependent Claim 23 words
The method according to claim 15 wherein LIF is added to the culture medium at a concentration of from 500 to 10,000 units/ml.

References Cited (Backward Citations)

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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) 1995-11-24 1996-05-25 1996-11-24 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1999-11-24 2000-05-25 2000-11-24 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2003-11-24 2004-05-25 2004-11-24 $7700.00 Window Closed / Lapsed

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