| Human mesenchymal stem cells (MSCs) are thought to be multilineage potential cells, which are present in bone marrow stromal, that can replicate as undifferentiated cells and that have the potential to differentiate to lineages of mesenchymal tissues or tissues from ectoderm, including bone, cartilage, skeletal muscle, tendon, ligament, corium, fat, marrow stroma and nerve. Therefore, MSCs could serve as seed cells constructing these tissues in Tissue Engineering.In order to find optimal seed cells applying to Tendon Tissue Engineering, MSCs were isolated from human fetal bone marrow aspirate, cultured and purified in this study. Their morphology, biological properties, telomerase activity, alkaline phosphatase, surface antigen and stroma protein were examined. These results suggested cultured MSCs be undifferentiated state. Multipotent differentiation ability of cultured MSCs was confirmed in nude mouse. Bone morphogenetic protein 12(BMP12) is an important regulator during tendon morphogenesis. Its cDNA including complete code sequences was cloned from full-term human placenta using RT-PCR and was linked correctly into multiple cloning region of pTARGET? a Eukaryocyte expression vector. pTARGET?- hBMP12 was transfected into MSCs by LipofectAMINE for inducing MSCs to commitment to tenocyte. The phenotypic features of transfected cells, different from parent MSCs, were similar to tenocytes, and these changes were determined in levels of morphology, mRNA and protein. Cell cycle analysis showed that transfected cells maintained active proliferation capacity resembling with parent MSCs.Stable and practical methods to isolate, culture expand MSCs and induce them to differentiate into tenocytes were established in this study. MSCs transfected bypTARGET? WBMP12 could serve as seed cells for Tendon Tissue Engineering. Moreover, this study provided academic and experimental basis for MSCs clinical applying to auto transplantation and regeneration of injured tendon.
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