Chemistry: molecular biology and microbiology – Spore forming or isolating process
Patent
1991-08-29
1995-05-23
Weimar, Elizabeth C.
Chemistry: molecular biology and microbiology
Spore forming or isolating process
424 937, 4352403, 800 2, 600 33, A61K 3700, C12N 500, C12N 1500, A61D 1900
Patent
active
054181593
DESCRIPTION:
BRIEF SUMMARY
The present invention relates generally to leukaemia inhibitory factor (LIF) from livestock species. More particularly, the present invention relates to the identification, cloning and structural characterisation of genes encoding LIF from livestock species. The present invention also relates to the use of LIF from livestock species in the enhancement of development of mammalian embryos and in maintaining ES cell lines in vitro.
LIF is a protein that has previously been cloned, produced and purified, in large quantities in purified recombinant form from both E. coli and yeast cells (International Patent Application No. PCT/AU88/00093.) LIF has been defined as a factor, the properties of which include: such as M1 cells, with associated differentiation of the leukaemic cells; and murine LIF or human LIF (defined in International Patent Application No. PCT/AU88/00093) for binding to specific cellular receptors on M1 cells or murine or human macrophages. In addition to the biological properties previously disclosed for murine and human LIF, LIF has been found to have the following additional properties: pluripotential phenotype of murine embryonic stem (ES) cell lines: D3 and EK-cs4 (derived from strain 129/SV blastocysts) and CBL63 and MD5 (derived from C57BL/6 blastocysts) as disclosed in International Application No. PCT/AU89/00330; the presence of LIF, to contribute to the tissues of chimaeric mice when re-introduced into the embryonic environment; ES (EK-cs1 and D3) and embryonic carcinoma (EC) (PCC3-3A, F9, PC13, P19 and MG2) cells; and competition with insulin, IGF-I, IGF-II, acidic and basic FGF, TGF.beta., TNF.alpha., TNF.beta., NGF, PDGF, EGF, IL-1, IL-2, IL-4, GM-CSF, G-CSF, Multi-CSF nor erythropoietin, but is in competition with murine and human LIF.
Accordingly, LIF is a potent hormone having utility in the general area of in vitro embryology, such as in maintaining ES cell lines and increasing the efficiency of embryo transfer and thus has important applications in the livestock industry. This is particularly apparent in the use of ES cells to provide a route for the generation of transgenic animals.
A major difficulty associated with present in vitro fertilisation (IVF) and embryo transfer (ET) programmes, particularly in humans, is the low success rate "achieved" on implantation of fertilised embryos. Currently, in human IVF programmes, the implantation rate may be as low as 10%, leading to the present practice of using up to four fertilised embryos in each treatment which, in turn, leads occasionally to multiple births. Accordingly, there is a need to improve the implantation rate in human IVF programmes. Similarly, in IVF and ET treatments in domestic animals such as sheep, cattle, pigs and goats, it is highly desirable for economic reasons to have as high as implantation rate as possible so as to reduce the numbers of fertilised embryos lost and unsuccessful treatment procedures performed.
In the development of a mammalian embryo, the fertilised egg passes through a number of stages including the morula and the blastocysts stages. In the blastocyst stage, the cells form an outer cell layer known as the trophectoderm (which is the precursor of the placenta) as well as an inner cells mass (from which the whole of the embryo proper is derived). The blastocyst is surrounded by the zona pellucida, which is subsequently lost when the blastocyst "hatches". The cells of the trophectoderm are then able to come into close contact with the wall of the uterus in the implantation stage. Prior to formation of the embryo proper by the inner cell mass by gastrulation, the whole cell mass may be referred to as "pre-embryo".
Although LIF from one species may be effective, for example in maintaining ES cell lines from a different or heterologous species, it may be preferable to develop homologous systems employing LIF and ES cell lines derived from the same species. It has now been found, in accordance with the present invention, that murine LIF DNA can be used to identify the LIF gene from a wide range of ma
REFERENCES:
patent: 5166065 (1992-11-01), Williams et al.
Robertson, et al. (1990) "In Vitro Models of the Maternal-Fetal Interface" in Molecular and Cellular Immunology of the Maternal-Fetal Interface, Wegmenn, et al., Oxford University Press, New York, 191-206.
Gearing, et al. (1987) "Molecular Cloning and Expression of cDNA Encoding a Murine Myeloid Leukaemia Inhibitory Factor (LIF)" EMBO J. 6, 3995-4002.
Smith, et al. (1988) "Inhibition of Pluripotential Embryonic Stem Cell Differentiation by Purified Polypeptides" Nature 336, 688-690.
Williams, et al. (1988) "Myeloid Leukaemia Inhibitory Factor Maintains the Developmental Potential of Embryonic Stem Cells" Nature 336, 684-687.
Fry et al (1992) Biology of Reproduction, 46, 470-474.
Gough Nicholas M.
Seamark Robert F.
Willson Tracey A.
Amrad Corporation Limited
Crouch Deborah
Weimar Elizabeth C.
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