Bone marrow is an important source of mesenchymal stem cells (MSCs), and a promising tool for cytotherapy. MSC utilization is limited by low cell yields obtained under standard isolation protocols. Herein, used bone marrow collection sets were evaluated as a valuable source of MSCs. Adherent cells washed from the collection sets were examined for widely accepted criteria defining MSCs. Significant numbers of cells (median 9million per set in passage 1) with colony-forming activity and high proliferative potential at low seeding densities were obtained. These cells were positive for essential MSC surface molecules (CD90, CD105, CD166, CD44, CD29) and negative for most haematopoietic and endothelial cell markers (CD45, CD34, CD11a, CD235a, HLA-DR, CD144). The cells were capable of differentiation along adipogenic, osteogenic and chondrogenic pathways. Washing out bone marrow collection sets may constitute a highly ethical source of MSCs for research purposes and may be utilized also in clinical applications.

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The effect of bovine seminal ribonuclease (BS RNase) on bone marrow cells in miniature pigs was studied. BS RNase at the concentration 20 and lOOug/ml preincubated 1 h with bone marrow cells from normal miniature pigs did not influence the formation of CFU-GM and BFU-E colonies cultivated in methylcellulose medium. The bone marrow cells kept in 4° C survived better in the presence of BS RNase than control cells cocultivated with bovine serum albumin (BSA). There was proved that pig bone marrow cells are able to bind the BS RNase on their surface without any morphological damage. For the experimental bone marrow transplantation in miniature pigs the whole body irradiation of 8G with a cobalt source was used. Two recipients from seven accepted semi-incompatible bone marrow cells preincubated 2 h with BS RNase (10mg/5xl0 8 cells

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