Cell Structure and Function
Online ISSN : 1347-3700
Print ISSN : 0386-7196
ISSN-L : 0386-7196
Isolation and Short-term Culture of Mouse Splenic Erythroblastic Islands
Yoshito SadahiraMasaharu MoriTetsuo Kimoto
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1990 年 15 巻 1 号 p. 59-65

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We isolated and cultured erythroblastic islands (EI) from the spleens of phlebotomized mice using a combination of collagenase digestion, unit gravity sedimentation, and Percoll density gradients separation. The isolated El were composed of surrounding erythroid cells and central stromal macrophages (Mφ), which were identified by Forssman antigen. While 60% of the erythroblasts incorporated bromodeoxyuridine, the Mφ did not. EI could be maintained on a plastic dish for a short period in the presence of erythropoietin.
Two hours later, the central Mφ spread well and bound to erythroblasts via cytoplasmic processes. One day later, erythropoietic activity on the Mφ surface continued, although their processes had retracted. Some EI showed synchronized expansion of erythroblasts and others showed differentiation to reticulocytes. Two days later, about 50% of the EI still showed erythropoietic activity and most erythroblasts differentiated to the orthochromatic stage. On the other hand, the Mφ secreted colony-stimulating activity during the culture. It was infrequently observed that erythroid and myeloid populations simultaneously expanded on a central Mφ. These results indicate that this El culture system is useful for studying interactions between the stomal Mφ and hematopoietic cells.
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© Japan Society for Cell Biology
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