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BACKGROUND: Recent?studies?show?that?mesenchymal?stem?cells?(MSCs)?have?immunomodulatory?properties.?They?suppress?the?immune?response?to?alloantigen?and?modify?the?proliferation?of?T?cells.?CD4+CD25+?regulatory?T?cells?have?strong?immunomodulatory?potential.?However,?little?is?known?about?the?effects?of?rat?MSCs?(rMSCs)?on?the?development?of?regulatory?T?cells. METHODS:MSCs?were?obtained?from?bone?marrow?of?male?Sprague-Dawley?rats,?and?co-cultured?with?CD3+?T?cells?from?allogeneic?spleen?cells.?The?proportion?of?CD4+CD25+?regulatory? T? cells? was? analyzed? by? lfow? cytometry.? To?further?conifrm?the?immunosuppressive?activity?of?rMSCs,?we?used?MTT?assay?and?lfow?cytometry?of?CD3+?T?cells?to?investigate?the?proliferative?responses?of?CD3+?T?cells?to?mitogenic?stimuli.?Enzyme-linked?immunosorbent?assay?was?performed?to?detect?alterations?of?the?cytokines?TNF-α,?TGF-β?and?IL-10. RESULTS:The?proliferation?of?CD3+?T?cells?decreased?when?co-cultured?with?rMSCs,?and?the?degree?of?inhibition?was?concentration-dependent.?The?percentage?of?CD4+CD25+?regulatory?T?cells?increased?when?CD3+?T?cells?were?co-cultured?with?different?concentrations?of?rMSCs.?The?levels?of?pro-inlfammatory?cytokine?(TNF-α)?decreased?while?anti-inlfammatory?(TGF-β,?IL-10)?cytokines?increased?in?mixed?lymphocyte?reaction. CONCLUSIONS:rMSCs?inhibit?allogeneic?T?cell?proliferation?in?mixed?cell?cultures.?This?immunosuppressive?effect?seems? to? be? mediated? by? inducing? the? generation? of?CD4+CD25+?regulatory?T?cells?and?soluble?factors.