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In order to identify the cDNAs responsible for hemopoietic supportive activity, expression of mRNAs in hemopoietic supportive bone marrow stromal cells (MS-5) and non-supportive stromal cells (MS-K) were compared by the cDNA subtraction method. A subtracted MS-5-cDNA library, which contains cDNA clones corresponding to mRNAs present in MS-5 cells but not in MS-K cells, was constructed. Screening of subtracted MS-5-cDNA library resulted in the isolation of some clones. Two of them, lipid binding protein (LBP) and haptoglobin (Hp), were expressed specifically in MS-5 cells but not in MS-K cells. The genes of LBP and Hp were subcloned into mammalian expression vector and transfected into hemopoietic non-supportive stromal cells line, MS-K. Then LBP-expressing stable transformants (MS-K-LBP) and Hp-expressing transformants (MS-K-Hp) were cloned. A rosette formation assay was carried out to investigate whether or not the LBP and Hp cause MS-K cells to adhere to hemopoietic cells. MS-K-LBP formed rosettes with hemopoietic cells as MS-5 cells, although the MS-K-Hp and normal MS-K cells did not form rosettes. These data indicate that LBP expressed in hemopoietic supportive stromal cells is partly responsible for the adhesion of hemopoietic stem cells to stromal cells.

cell structure and function

Gene Expression of Lipid Binding Protein Transferred the Ability of Specific Attachment of Hemopoietic Cells to Non-supportive Stromal Cell Line, MS-K.