A Quantitative Assay for Stroma Dependent Hemopoiesis
Hemopoiesis is strictly dependent upon the self renewal of pluripotent stem cells in the bone marrow. A small fraction of these cells differentiates in a series of steps and continuously supplies mature cells to the myeloid and immune systems. Ultrastructural studies of the bone marrow have revealed close physical associations of stromal cells with hemopoietic stem and progenitor cells [1, 2]. Continuous hemopoiesis and long term maintenance of stem cells in vitro can be obtained only in coculture with bone marrow stromal cells . Functional significance has been ascribed mostly to stromal-adipose cells . A number of laboratories have reported on the establishment of continuous cell lines from mouse bone marrow stroma. These cell lines have been characterized morphologically and functionally and some were shown to support both myelopoiesis and pre-B lymphopoiesis in long term cultures [5, 6, 7, 8, 9]. Although these cell lines have significantly simplified the model systems used for the study of hemopoiesis in vitro, the cultures are very complex and are hard to use when one is trying to evaluate the molecular basis of the mechanisms that control the process. Extracellular matrix components were proposed to be involved in the regulation of hemopoiesis. It was found that stromal cells deposit high amounts of extracellular matrix components, some of which were detected at attachment sites between marrow derived adherent cells and developing granulocytes and macrophages [10, 11]. So far the exact role of the extracellular matrix is unknown.
KeywordsMigration Rubber Fluorescein FITC Glycosaminoglycan
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