Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1989 Oct;9(10):4541–4544. doi: 10.1128/mcb.9.10.4541

Mock retroviral infection alters the developmental potential of murine bone marrow stem cells.

D Dumenil 1, H Jacquemin-Sablon 1, H Neel 1, E Frindel 1, F Dautry 1
PMCID: PMC362540  PMID: 2573833

Abstract

Retroviral vectors were used to introduce an activated ras gene into murine pluripotent hemopoietic stem cells. We attempted to reconstitute the hemopoietic system of lethally irradiated mice with isolated spleen colonies obtained in vivo after injection of infected bone marrow cells. Spleen colonies derived from infected bone marrow were inefficient in promoting long-term survival of irradiated hosts. This loss of reconstitutive capacity of spleen colonies was not due to the retroviral infection per se but to the in vitro culture of spleen colony precursors. Incubation for 24 h in the presence of fetal calf serum and interleukin-3 without virus-producing cells was sufficient to abolish completely the reconstitutive capacity of spleen colonies while maintaining both self-renewal and pluripotential capacities of spleen colony precursors. These results show that the in vitro manipulation of stem cells that is included in current protocols for retroviral infection can modify the developmental potential of these cells. This finding clearly indicates that the use of retroviral vectors can introduce a bias in the analysis of hemopoiesis.

Full text

PDF
4541

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  2. Bertoncello I., Hodgson G. S., Bradley T. R. Multiparameter analysis of transplantable hemopoietic stem cells. II. Stem cells of long-term bone marrow-reconstituted recipients. Exp Hematol. 1988 May;16(4):245–249. [PubMed] [Google Scholar]
  3. Cepko C. L., Roberts B. E., Mulligan R. C. Construction and applications of a highly transmissible murine retrovirus shuttle vector. Cell. 1984 Jul;37(3):1053–1062. doi: 10.1016/0092-8674(84)90440-9. [DOI] [PubMed] [Google Scholar]
  4. Dick J. E., Magli M. C., Huszar D., Phillips R. A., Bernstein A. Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/Wv mice. Cell. 1985 Aug;42(1):71–79. doi: 10.1016/s0092-8674(85)80102-1. [DOI] [PubMed] [Google Scholar]
  5. Eglitis M. A., Kantoff P., Gilboa E., Anderson W. F. Gene expression in mice after high efficiency retroviral-mediated gene transfer. Science. 1985 Dec 20;230(4732):1395–1398. doi: 10.1126/science.2999985. [DOI] [PubMed] [Google Scholar]
  6. Hawley R. G., Covarrubias L., Hawley T., Mintz B. Handicapped retroviral vectors efficiently transduce foreign genes into hematopoietic stem cells. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2406–2410. doi: 10.1073/pnas.84.8.2406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hodgson G. S., Bradley T. R. Properties of haematopoietic stem cells surviving 5-fluorouracil treatment: evidence for a pre-CFU-S cell? Nature. 1979 Oct 4;281(5730):381–382. doi: 10.1038/281381a0. [DOI] [PubMed] [Google Scholar]
  8. Hodgson G. S., Bradley T. R., Radley J. M. The organization of hemopoietic tissue as inferred from the effects of 5-fluorouracil. Exp Hematol. 1982 Jan;10(1):26–35. [PubMed] [Google Scholar]
  9. Karlsson S., Bodine D. M., Perry L., Papayannopoulou T., Nienhuis A. W. Expression of the human beta-globin gene following retroviral-mediated transfer into multipotential hematopoietic progenitors of mice. Proc Natl Acad Sci U S A. 1988 Aug;85(16):6062–6066. doi: 10.1073/pnas.85.16.6062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Keller G., Paige C., Gilboa E., Wagner E. F. Expression of a foreign gene in myeloid and lymphoid cells derived from multipotent haematopoietic precursors. Nature. 1985 Nov 14;318(6042):149–154. doi: 10.1038/318149a0. [DOI] [PubMed] [Google Scholar]
  11. Lemischka I. R., Raulet D. H., Mulligan R. C. Developmental potential and dynamic behavior of hematopoietic stem cells. Cell. 1986 Jun 20;45(6):917–927. doi: 10.1016/0092-8674(86)90566-0. [DOI] [PubMed] [Google Scholar]
  12. Magli M. C., Iscove N. N., Odartchenko N. Transient nature of early haematopoietic spleen colonies. Nature. 1982 Feb 11;295(5849):527–529. doi: 10.1038/295527a0. [DOI] [PubMed] [Google Scholar]
  13. Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
  14. McCoy M. S., Bargmann C. I., Weinberg R. A. Human colon carcinoma Ki-ras2 oncogene and its corresponding proto-oncogene. Mol Cell Biol. 1984 Aug;4(8):1577–1582. doi: 10.1128/mcb.4.8.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Micklem H. S., Lennon J. E., Ansell J. D., Gray R. A. Numbers and dispersion of repopulating hematopoietic cell clones in radiation chimeras as functions of injected cell dose. Exp Hematol. 1987 Mar;15(3):251–257. [PubMed] [Google Scholar]
  16. Mulder A. H., Visser J. W. Separation and functional analysis of bone marrow cells separated by rhodamine-123 fluorescence. Exp Hematol. 1987 Jan;15(1):99–104. [PubMed] [Google Scholar]
  17. Reincke U., Rosenblatt M., Hellman S. An in vitro clonal assay of adherent stem cells (ASC) in mouse marrow. J Cell Physiol. 1984 Nov;121(2):275–283. doi: 10.1002/jcp.1041210203. [DOI] [PubMed] [Google Scholar]
  18. Shibagaki T., Inoue T., Kubota N., Kanisawa M. Fraction of pluripotent hemopoietic stem cells in DNA synthesis varies with generation age. Exp Hematol. 1986 Sep;14(8):794–797. [PubMed] [Google Scholar]
  19. Snodgrass R., Keller G. Clonal fluctuation within the haematopoietic system of mice reconstituted with retrovirus-infected stem cells. EMBO J. 1987 Dec 20;6(13):3955–3960. doi: 10.1002/j.1460-2075.1987.tb02737.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spangrude G. J., Heimfeld S., Weissman I. L. Purification and characterization of mouse hematopoietic stem cells. Science. 1988 Jul 1;241(4861):58–62. doi: 10.1126/science.2898810. [DOI] [PubMed] [Google Scholar]
  21. Spooncer E., Lord B. I., Dexter T. M. Defective ability to self-renew in vitro of highly purified primitive haematopoietic cells. Nature. 1985 Jul 4;316(6023):62–64. doi: 10.1038/316062a0. [DOI] [PubMed] [Google Scholar]
  22. TILL J. E., MCCULLOCH E. A., SIMINOVITCH L. A STOCHASTIC MODEL OF STEM CELL PROLIFERATION, BASED ON THE GROWTH OF SPLEEN COLONY-FORMING CELLS. Proc Natl Acad Sci U S A. 1964 Jan;51:29–36. doi: 10.1073/pnas.51.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TILL J. E., McCULLOCH E. A. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat Res. 1961 Feb;14:213–222. [PubMed] [Google Scholar]
  24. Williams D. A., Lemischka I. R., Nathan D. G., Mulligan R. C. Introduction of new genetic material into pluripotent haematopoietic stem cells of the mouse. Nature. 1984 Aug 9;310(5977):476–480. doi: 10.1038/310476a0. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES