1974 Volume 15 Issue 9 Pages 1002-1010
The proliferation and differentiation of bone marrow hematopoietic cells in normal children and children with acute leukemia and idiopathic aplastic anemia were studied in vitro.
In vitro diffusion chamber technic, described by Sumner et al and Golde et al, was employed as the method of the cell culture.
McCoy's 5A Medium supplemented with 15% fetal calf serum and antibiotics, were used without addition of exogenous granulocyte stimulating substances.
The culture apparatus contained the bone marrow cells (2-6×106) were incubated at 37°C in a humidified environment with 7.5% CO2 in air.
The cultures were harvested up to 14 days and sequential observations were done on viable cell counts, differential counts of May-Grunwald-Giemsa stained cells, and radioautographs, prepared after incubation of the cells with 1μC/ml of 3H-Thymidine for 1 hour.
Briefly, the results were as follows:
1) In the culture of normal bone marrow, various maturational stages of granulocytes were found after the 10-14th day, and small numbers of lymphocyts, erythroblastes and rare megakaryocytes were found. The macrophages increased gradually.
The viable cell counts were falling until 3-5th day, followed by a rise or slight decrease toward the 14th day.
3H-Thymidine up-take rate (labelling indices) paralled to the variations of cell counts.
2) The growth of leukemic cells from children with acute lymphoblastic leukemia showed maturation arrest. The cells of acute myeloblastic leukemia showed partially cytoplasmic changes with irregular shape and bulky vacuoles.
3) In idiopathic aplastic anemia, although small numbers of mature granulocytes were presented, lymphoid cells and macrophages were predominant.
