Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Effects of a preformed extracellular matrix on long-term serum-free bone marrow culture

Summary

The extracellular matrix (ECM) produced by the stromal layer plays a key role in the regulation of commitment and differentiation of hematopoietic cells. Long-term bone marrow culture (LTBMC) allows analysis of the stromal microenvironment. Recently, serum-free LTBMC has been described, but the formation of a classical adherent layer was never observed under these conditions. We have evaluated the effect(s) of a chemically well defined ECM on serum-free and serum-dependent LTBMC. In serum-dependent cultures ECM did not induce a significant increase of hematopoiesis. In serum-free conditions, a marked improvement of hematopoiesis was observed, both in terms of CFU-GM and BFU-E yield and in duration of cultures. A confluent stromal layer was observed only in the presence of ECM. The present results indicate that the addition of ECM to serum-free cultures provides a standardized culture condition, while improving progenitor cell recovery and allowing formation of a confluent stromal layer. Moreover, ECM+ LTBMC may provide a model to study the effect(s) of adhesive proteins and hematopoietic growth factors normally present in serum.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Aye MT, Niho Y, Till JE, McCulloch EA (1974) Studies of leukemic cell populations in culture. Blood 44: 205–219

  2. 2.

    Chang J, Morgenstern GR, Coutinho LH, Scarffe JH, Carr T, Deakin DP, Testa NG, Dexter TM (1989) The use of bone marrow cells grown in long-term culture for autologous bone marrow transplantation in acute myeloid leukaemia: an update. Bone Marrow Transplant 4: 5–9

  3. 3.

    Coulombel L, Eaves AC, Eaves CJ (1983) Enzymatic treatment of long-term human marrow cultures reveals the preferential location of primitive hemopoietic progenitors in the adherent layer. Blood 62: 291–297

  4. 4.

    Coulombel L, Kalousek DK, Eaves CJ, Gupta CM, Eaves AC (1983) Long-term marrow culture reveals chromosomally normal hemopoietic progenitor cells in patients with Philadelphia chromosome-positive chronic myelogenous leukemia. N Engl J Med 308: 1493–1498

  5. 5.

    Coulombel L, Vuillet MH, Leroy C, Tchernia G (1988) Lineage and stage-specific adhesion of human hematopoietic progenitor cells to extracellular matrices from marrow fibroblasts. Blood 71: 329–334

  6. 6.

    Drouet X, Douay L, Giarratana MC, Baillou C, Gorin NC, Salmon C, Najman A (1989) Human liquid bone marrow culture in serum-free medium. Br J Haematol 73: 143–147

  7. 7.

    Eliason JF (1986) Granulocyte-macrophage colony formation in serum-free culture: effects of purified colony stimulating factors and modulation by hydrocortisone. J Cell Physiol 128: 231

  8. 8.

    Gabbianelli M, Pelosi E, Bassano E, Labbaye C, Petti S, Tritarelli E, Miller BA, Valtieri M, Testa U, Peschle C (1989) Granulocyte-macrophage colony-stimulating factor reactivates fetal hemoglobin synthesis in erythroblast clones from normal adults. Blood 74: 2657–2667

  9. 9.

    Gartner S, Kaplan HS (1980) Long-term culture of human bone marrow cells. Proc Natl Acad Sci USA 77: 4756–4759

  10. 10.

    Gordon MY, Dowding CR, Riley GP, Goldman JM, Greaves MF (1987) Altered adhesive interactions with marrow stroma of haematopoietic progenitor cells in chronic myeloid leukaemia. Nature 328: 342–344

  11. 11.

    Gordon MY, Riley GP, Watt SM, Greaves MF (1987) Compartmentalization of a haematopoietic growth factors (GM-CSF) by glycosaminoglycans in the bone marrow microenvironment. Nature 326: 403–405

  12. 12.

    Heard JM, Fichelson S, Varet B (1982) Role of colony stimulating activity in murine long-term bone marrow cultures: evidence for its production and consumption by the adherent cells. Blood 59: 761–767

  13. 13.

    Lichtman MA (1981) The ultrastructure of the hemopoietic environment of the marrow: a review. Exp Hematol 9: 391–410

  14. 14.

    Liesveld JL, Winslow JM, Kempski MC, Ryan DH, Brennan JK, Abboud CN (1991) Adhesive interactions of normal and leukemic CD 34+ myeloid progenitors: role of marrow stroma, fibroblast and cytomatrix components. Exp Hematol 19: 63–70

  15. 15.

    Merghenthaler HG, Dormer P (1990) Hemopoiesis in human micro long-term bone marrow culture with preformed extracellular matrix. Haematologica (Pavia) 75: 12–16

  16. 16.

    Ming DA, Douay L, Barbu V, Fabrega S, Allieri MA, Drouet X, Deloucs J, Giarratana MC, Oszhain H, van den Akker J, Vanhaeke D, Gorin NC (1991) Serum-free liquid marrow culture in patients with acute lymphoblastic leukaemia: a potential application to purge marrow for autologous transplantation. Br J Haematol 78: 42–47

  17. 17.

    Roberts RA, Spooncer E, Parkinson EK, Lord BI, Allen TD, Dexter TM (1987) Metabolically inactive 3T3 cells can substitute for marrow stromal cells to promote the proliferation and development of multipotent hemopoietic stem cells. J Cell Physiol 132: 203–214

  18. 18.

    Sadovnikova EY, Deryugina EI, Drize NJ, Chertkov JL (1991) Induction of hematopoietic microenvironment by the extra-cellular matrix from long-term bone marrow cultures. Ann Hematol 62: 160–164

  19. 19.

    Spooncer E, Gallagher JT, Krizsa F, Dexter TM (1983) Regulation of hemopoiesis in long-term bone marrow cultures. IV. Glycosaminoglycan synthesis and the stimulation of hemopoiesis by beta-D-xylosides. J Cell Biol 96: 510–514

  20. 20.

    Strobel ES, Strobel HG, Bross KJ, Winterhalter B, Fiebig HH, Schildge JU, Lohr GW (1989) Effects of human bone marrow stroma on the growth of human tumor cells. Cancer Res 49: 1001–1007

Download references

Author information

Correspondence to G. Leone.

Additional information

This work was supported by an AIRC grant and the Program “Terapia dei Tumori”,Istituto Superiore di Sanità, Rome

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Teofili, L., Sargiacomo, M., Iovino, M.S. et al. Effects of a preformed extracellular matrix on long-term serum-free bone marrow culture. Ann Hematol 65, 22–25 (1992). https://doi.org/10.1007/BF01715121

Download citation

Key words

  • Extracellular matrix
  • Human long-term bone marrow culture