Ex-Vivo Treatment of Graft by Long-Term Culture
To establish human long-term bone marrow cultures (LTBMC), 2 X 107 nucleated bone marrow cells are suspended in 10 ml of Iscove’s medium containing 10% (vol: vol) fetal calf serum, 10% (vol: vol) horse serum and 5x101M hydrocortisone and added to flat bottomed plastic flasks which have been gassed with a mixture of 5% C02 in air. The cultures are maintained at 33 °C and at weekly intervals half the medium is removed and replaced with fresh medium. The cells in the overlying medium contain differentiated haematopoietic cells, mainly macrophages, neutrophils and progenitor cells, which can be assayed in soft gels for their ability to form colonies. Haematopoiesis and the production of progenitor cells in LTBMC can be maintained for 8 weeks or more .
KeywordsAcute Myeloid Leukaemia Haematopoietic Stem Cell Leukaemic Cell Haematopoietic Cell Acute Myeloid Leukaemia Patient
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- 1.Allen TD, Dexter TM, Simmons PJ (1990) Marrow biology and stem cells. In: Dexter TM, Garland JM, Testa NG (eds) Colony stimulating factors: cellular and molecular biology. Dekker, New York, pp 1–38 (Haematology, vol 9 )Google Scholar
- 3.Chang J, Morgenstern GR, Coutinho LH, Scarffe J, Carr T, Deakin DP, Testa NG, Dexter TM (1989 a) The use of bone marrow cells grown in long-term culture for autologous BMT in acute myeloid leukaemia: an update. Bone Marrow Transplant 4: 5–9Google Scholar
- 7.Coutinho LH, Will A, Chang J, Testa NG, Dexter TM (1987) Long-term bone marrow cultures in autologous transplantation: an in-vitro biological system for purging leukaemic cells. In: Baum SJ, Santos GW, Takaku F (eds) Experimental hematology today 1987. Recent advances and future directions in bone marrow transplantation. Springer, Berlin Heidelberg New York, pp 106–113Google Scholar
- 10.Gordon MY, Riley GP, Watt SM, Greaves MF (1987a) Compartmentalization of a haematopoietic growth factor ( GM-CSF) by glycosaminoglycans in the bone marrow environment. Nature 326: 403–405Google Scholar
- 12.McCulloch EA, Minden MD, Miyauchi J, Kelleher CA, Wang C (1988) Stem cell renewal and differentiation in acute myeloid leukaemia. J Cell Sci [Suppl 10]: 267–281Google Scholar
- 14.Winton EF, Colenda KW (1987) The use of long-term human marrow cultures to demonstrate progenitor cell precursors in marrow treated with 4-hydroperoxycyclo- phosphamide. Exp Hematol 13: 710–714Google Scholar