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The developed Leydig cells-conditioned medium (LCM) contains bioactive materials secreted by Leydig cells in vitro .  LCM was used to evaluate the ability of bone marrow mesenchymal stem cells differentiation. Bone marrow mesenchymal stem cells (1x 10 6 cell/ml) were cultured in : 1) DMEM supplemented with 10% NBCS as a control (M), 2) M supplemented with 10 ng/ml testosterone; 3) M supplemented with 50%  LCM ; 4) M supplemented with 50% LCM and 2.5 IU/ml hCG. Bone marrow mesenchymal stem cells that were cultured with  LCM has a positive reaction (57.4%) to histochemistry staining 3β-HSD and produced 1.87 ng/ml testosterone. Supplementation of hCG to LCM  increased the positive number of Leydig cells and testosterone production by 74.6% and 12.33 ng/ml (P<0.05). It can be concluded that Leydig cells-conditioned medium can support differentiation of bone marrow mesenchymal stem cells into Leydig cells.

Biological Analysis of Leydig Cells-Conditioned Medium To Support Rat Bone Marrow Mesenchymal Stem Cells Differentiation