Isolation and Short-term Culture of Mouse Splenic Erythroblastic Islands

Cell Structure and Function

Yoshito Sadahira¹  Masaharu Mori²  Tetsuo Kimoto¹ 

[1] Department of Pathology, Kawasaki Medical School;Department of Pathology, Okayama University Medical School

关键词: erythropoiesis;    meylopoiesis;    macrophage;    erythropoietin;    colony-stimulating factor;   

DOI  :  10.1247/csf.15.59

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(21)Cited-By(3)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.

【 授权许可】

Unknown