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J. Biol. Chem., Vol. 276, Issue 12, 9189-9198, March 23, 2001
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From The Beatson Institute for Cancer Research, Cancer Research
Campaign Beatson Laboratories, Garscube Estate, Switchback Road,
Bearsden, Glasgow G61 1BD, United Kingdom
The difficulties associated with
studying molecular mechanisms important in hemopoietic stem cell (HSC)
function such as the problems of purifying homogeneous stem cell
populations, have prompted us to adapt the murine ES cell system as an
in vitro model of HSC generation and function. We now
report that careful analysis of the time course of HSC generation in
differentiating ES cells allows them to be used as a source of known
and novel hemopoietic gene products. We have generated a subtracted
library using cDNA from ES cells collected just prior to and just
following the emergence of HSCs. Analysis of this library shows it to
be a rich source of known hemopoietic and hemopoietic related gene products with 44% of identifiable cDNAs falling into these camps. We have demonstrated the value of this system as a source of novel genes of relevance to HSC function by characterizing a novel membrane protein encoding cDNA that is preferentially expressed in primitive hemopoietic cells. Intriguingly, further analysis of the known components of the subtracted library is suggestive of erythroid preconditioning of the ES cell-derived HSC. We have used dot-blot and
in situ analysis to indicate that this erythroid
preconditioning is probably restricted to primitive but not definitive
HSC.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF009781.
Differentiating Embryonal Stem Cells Are a Rich Source
of Haemopoietic Gene Products and Suggest Erythroid Preconditioning of
Primitive Haemopoietic Stem Cells*
,
,, and
*
This work was supported by grants from the Medical Research
Council and the Leukemia Research Fund. Work at the Beatson Institute was supported by grants from the Cancer Research Campaign.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Present address: Regulation of Cell Growth Laboratory, Bldg. 560, Rm. 22-45, NCI-Frederick Cancer Research and Development Center,
Frederick, MD 21702.
§
Present address: EiRx Therapeutics Ltd., Bldg. 2800, Kinsale Road,
Cork, Eire, United Kingdom.
¶
Present address: Academic Unit of Obstetrics, Gynaecology and
Reproductive Healthcare, St. Mary's Hospital, Manchester, M13 0JH
United Kingdom.
Present address: Patterson Institute for Cancer Research,
Wilmslow Road, Manchester, M20 4BX United Kingdom.
**
Present address: John Hughes Bennett Laboratory, Dept. of Oncology,
Western General Hospital, Edinburgh, EH4 2XU, United Kingdom.
Present address: Biological Sciences, University of Durham,
South Rd., Durham DH1 3LE, United Kingdom.
§§
To whom correspondence should be addressed: Beatson Institute for
Cancer Research, Cancer Research Campaign Beatson Laboratories, Garscube Estate, Switchback Road, Bearsden, Glasgow G61 1BD, United Kingdom. Tel.: 44-141-330-3982; Fax: 44-141-942-6521; E-mail: g.graham@beatson.gla.ac.uk.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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