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Stem Cells, Vol. 15, No. 3, 229-236, May 1997
© 1997 AlphaMed Press

Adhesion of Thymocytes to Bone Marrow Stromal Cells: Regulation by bFGF and IFN-{gamma}

Mira Barda-Saada, An-Sheng Zhanga, Dov Ziporib, Leon A. Rozenszajna

a Department of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel;
b Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot, Israel

Key Words. T cell-bone marrow stroma cell adhesion • Microenvironment • Fluorimetric cell quantitation • Calcein-AM • FDA • bFGF • IFN-{gamma}

Dr. Dov Zipori, Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot 76100, Israel.

We recently reported on selective interactions between immature T cell subpopulations and bone marrow (BM) stromal cells. To further study this process, we first examined the efficacy of methods estimating cell-cell adhesion and then investigated the effects of cytokines on thymocyte-stroma associations. Techniques based on the use of the fluorochromes calcein-acetomethylester (calcein-AM) and fluorescein diacetate (FDA) were studied and compared to regular cell counting methods. With calcein-AM labeling, the retention time was relatively long, while with FDA labeling, there was a rapid cellular efflux. Using calcein-AM, we developed an accurate quantitative fluorometric assay for determining the adherence of thymocytes to a BM stromal cell line (MBA-13). A maximal fraction of about 29% thymocytes was found to adhere to confluent MBA-13 cell layers after four to six h of coculture. Whereas interleukin 1 did not change the rate of adhesion of thymocytes to the stroma, interferon-{gamma} (IFN-{gamma}) significantly increased adhesion. Basic fibroblast growth factor (bFGF) had a dose-dependent biphasic effect on thymocyte adhesion, and a greater fraction of double negative thymocytes adhered to stroma pretreated with bFGF. Taken together, these results suggest that IFN-{gamma} and bFGF modulate T cells-BM stromal cell adhesion.




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