Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1982 May 1;155(5):1370–1384. doi: 10.1084/jem.155.5.1370

Proliferation and differentiation of hematopoietic stem cells in long- term cultures of adult hamster spleen

PMCID: PMC2186680  PMID: 7069371

Abstract

We have found that in liquid cultures of spleen cells of adult Syrian hamsters of the F1D strain, the hematopoietic microenvironment is adequate to sustain proliferation of splenic stem cells for periods of greater than 4 mo, and permits granulocytic, monocytic, and megakaryocytic differentiation without secondary repopulation or addition of exogenous growth factors to the basic medium of RPMI 1640 plus 20% horse serum. Intimate topographical relations are established between spleen stromal cells and hematopoietic cell components of the culture is adherent "cell-producing" islets. Some of these islets are associated with multiple hematopoietic cell types such as myeloid, monocytic, and megakaryocytic cells. Other islets are associated with a single cell type such as megakaryocytes, which suggests a limited potential of some adherent stromal cells to direct the differentiation of precursor cells. Cultures of this type provide a simple and convenient model for investigation of the mechanisms controlling differentiation of hematopoietic stem cells, not only for granulocytic and monocytic cells, but for megakaryocytic cells as well.

Full Text

The Full Text of this article is available as a PDF (3.4 MB).

Selected References

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

  1. Allen T. D., Dexter T. M. Cellular interrelationships during in vitro granulopoiesis. Differentiation. 1976 Oct 7;6(3):191–194. doi: 10.1111/j.1432-0436.1976.tb01486.x. [DOI] [PubMed] [Google Scholar]
  2. Burstein S. A., Adamson J. W., Thorning D., Harker L. A. Characteristics of murine megakaryocytic colonies in vitro. Blood. 1979 Jul;54(1):169–179. [PubMed] [Google Scholar]
  3. Cline M. J., Golde D. W. Controlling the production of blood cells. Blood. 1979 Jan;53(1):157–165. [PubMed] [Google Scholar]
  4. Curry J. L., Trentin J. J. Hemopoietic spleen colony studies. I. Growth and differentiation. Dev Biol. 1967 May;15(5):395–413. doi: 10.1016/0012-1606(67)90034-6. [DOI] [PubMed] [Google Scholar]
  5. Dexter T. M., Allen T. D., Lajtha L. G. Conditions controlling the proliferation of haemopoietic stem cells in vitro. J Cell Physiol. 1977 Jun;91(3):335–344. doi: 10.1002/jcp.1040910303. [DOI] [PubMed] [Google Scholar]
  6. Dexter T. M., Testa N. G. Differentiation and proliferation of hemopoietic cells in culture. Methods Cell Biol. 1976;14:387–405. doi: 10.1016/s0091-679x(08)60498-7. [DOI] [PubMed] [Google Scholar]
  7. Dexter T. M., Testa N. G. In vitro methods in haemopoiesis and lymphopoiesis. J Immunol Methods. 1980;38(3-4):177–190. doi: 10.1016/0022-1759(80)90266-5. [DOI] [PubMed] [Google Scholar]
  8. Golde D. W., Faille A., Cline M. J. Induction of splenic granulopoiesis in vitro. Proc Soc Exp Biol Med. 1976 Sep;152(4):544–548. doi: 10.3181/00379727-152-39436. [DOI] [PubMed] [Google Scholar]
  9. Guilbert L. J., Iscove N. N. Partial replacement of serum by selenite, transferrin, albumin and lecithin in haemopoietic cell cultures. Nature. 1976 Oct 14;263(5578):594–595. doi: 10.1038/263594a0. [DOI] [PubMed] [Google Scholar]
  10. Haley J. E., Tjio J. H., Smith W. W., Brecher G. Hematopoietic differentiative properties of murine spleen implanted in the omenta of irradiated and nonirradiated hosts. Exp Hematol. 1975 Jun;3(3):187–196. [PubMed] [Google Scholar]
  11. Johnson G. R., Metcalf D. Pure and mixed erythroid colony formation in vitro stimulated by spleen conditioned medium with no detectable erythropoietin. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3879–3882. doi: 10.1073/pnas.74.9.3879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KARNOVSKY M. J., ROOTS L. A "DIRECT-COLORING" THIOCHOLINE METHOD FOR CHOLINESTERASES. J Histochem Cytochem. 1964 Mar;12:219–221. doi: 10.1177/12.3.219. [DOI] [PubMed] [Google Scholar]
  13. Korn A. P., Henkelman R. M., Ottensmeyer F. P., Till J. E. Investigations of a stochastic model of haemopoiesis. Exp Hematol. 1973;1(6):362–375. [PubMed] [Google Scholar]
  14. La Pushin R. W., Trentin J. J. Identification of distinctive stromal elements in erythroid and neutrophil granuloid spleen colonies: light and electron microscopic study. Exp Hematol. 1977 Nov;5(6):505–522. [PubMed] [Google Scholar]
  15. Lidbeck J., Månson J. C. Staining of granulopoietic agar colonies with a modified Papanicolaou technique. Exp Hematol. 1979 Mar;7(3):166–169. [PubMed] [Google Scholar]
  16. Metcalf D., Johnson G. R., Mandel T. E. Colony formation in agar by multipotential hemopoietic cells. J Cell Physiol. 1979 Feb;98(2):401–420. doi: 10.1002/jcp.1040980216. [DOI] [PubMed] [Google Scholar]
  17. Nussenzweig M. C., Steinman R. M. Contribution of dendritic cells to stimulation of the murine syngeneic mixed leukocyte reaction. J Exp Med. 1980 May 1;151(5):1196–1212. doi: 10.1084/jem.151.5.1196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Salmon S. E., Buick R. N. Preparation of permanent slides of intact soft-agar colony cultures of hematopoietic and tumor stem cells. Cancer Res. 1979 Mar;39(3):1133–1136. [PubMed] [Google Scholar]
  19. Steinman R. M., Adams J. C., Cohn Z. A. Identification of a novel cell type in peripheral lymphoid organs of mice. IV. Identification and distribution in mouse spleen. J Exp Med. 1975 Apr 1;141(4):804–820. [PMC free article] [PubMed] [Google Scholar]
  20. Steinman R. M., Cohn Z. A. Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J Exp Med. 1973 May 1;137(5):1142–1162. doi: 10.1084/jem.137.5.1142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Steinman R. M., Witmer M. D. Lymphoid dendritic cells are potent stimulators of the primary mixed leukocyte reaction in mice. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5132–5136. doi: 10.1073/pnas.75.10.5132. [DOI] [PMC free article] [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. Trentin J. J. Determination of bone marrow stem cell differentiation by stromal hemopoietic inductive microenvironments (HIM). Am J Pathol. 1971 Dec;65(3):621–628. [PMC free article] [PubMed] [Google Scholar]
  25. Trentin J. J. Hemopoietic microenvironments. Transplant Proc. 1978 Mar;10(1):77–82. [PubMed] [Google Scholar]
  26. Wolf N. S., Trentin J. J. Hemopoietic colony studies. V. Effect of hemopoietic organ stroma on differentiation of pluripotent stem cells. J Exp Med. 1968 Jan 1;127(1):205–214. doi: 10.1084/jem.127.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wright E. G., Lord B. I., Dexter T. M., Lajtha L. G. Mechanisms of haemopoietic stem cell proliferation control. Blood Cells. 1979 Jun 15;5(2):247–258. [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES