A fibronectin matrix is required for differentiation of murine erythroleukemia cells into reticulocytes.
A fibronectin matrix is required for differentiation of murine erythroleukemia cells into reticulocytes.
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DOI：
10.1083/jcb.105.6.3105
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发表时间：
1987-12
期刊：
影响因子：
--
通讯作者：
Lodish HF
中科院分区：
文献类型：
--
作者：
Patel VP;Lodish HF
小鼠红白血病（MEL）细胞的红系分化是更广泛的，当细胞连接到纤连蛋白包被的盘比悬浮培养。在悬浮培养4天诱导的细胞在晚期成红细胞阶段被捕，并且不经历去核。细胞在悬浮液中孵育超过4天导致裂解。与此相反，DMSO诱导的细胞在纤连蛋白包被的培养皿7天分化成去核细胞，网织红细胞和红细胞。如通过定量免疫印迹法所测定的，悬浮培养物中诱导的细胞积累了红细胞中存在的主要红细胞膜蛋白带3量的约33%，而纤连蛋白包被的培养皿上诱导的细胞积累了红细胞中存在的量的80-100%。悬浮诱导的细胞和在纤连蛋白包被的培养皿上诱导的细胞都积累了红细胞中存在的血影蛋白和锚蛋白量的约90%。如MEL细胞去核过程中免疫荧光显微镜所示，带3和锚蛋白都被隔离在新生网织红细胞的胞质片段中。由于表面纤连蛋白受体的丢失，去核和晚期细胞从纤连蛋白基质中分离;这模拟了网织红细胞从骨髓基质正常释放到血液中。因此，纤连蛋白基质提供了一个允许的微环境，其中红系前体细胞驻留，增殖，迁移和表达其正常分化程序。
Erythroid differentiation of murine erythroleukemia (MEL) cells is far more extensive when the cells are attached to fibronectin-coated dishes than in suspension culture. Cells induced in suspension culture for 4 d become arrested at a late erythroblast stage and do not undergo enucleation. Incubation of cells in suspension beyond 4 d results in lysis. In contrast, cells induced by DMSO on fibronectin-coated dishes for 7 d differentiate into enucleating cells, reticulocytes, and erythrocytes. As determined by quantitative immunoblotting, cells induced in suspension culture accumulate approximately 33% of the amount of the major erythroid membrane protein Band 3 present in erythrocyte, whereas cells induced on fibronectin-coated dishes accumulate 80-100% of the amount present in erythrocytes. Both suspension-induced cells and cells induced on fibronectin-coated dishes accumulate approximately 90% of the amount of spectrin and ankyrin present in erythrocytes. As revealed by immunofluorescence microscopy during enucleation of MEL cells, both Band 3 and ankyrin are sequestered in the cytoplasmic fragment of the emerging reticulocyte. Enucleated and later-stage cells detach from the fibronectin matrix, due to the loss of the surface fibronectin receptor; this mimics the normal release of reticulocytes from the matrix of the bone marrow into the blood. Thus a fibronectin matrix provides a permissive microenvironment within which erythroid precursor cells reside, proliferate, migrate, and express their normal differentiation program.