Patruno Marco, Gomiero Chiara, Sacchetto Roberta, Topel Ohad, Negro Alessandro, Martinello Tiziana
Department of Comparative Biomedicine and Food Science, University of Padova, Padova, Italy.
VTH - Koret School of Veterinary Medicine, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Jerusalem, Israel.
Vet Res Commun. 2017 Sep;41(3):211-217. doi: 10.1007/s11259-017-9692-y. Epub 2017 Jun 6.
The Tat protein is able to translocate through the plasma membrane and when it is fused with other peptides may acts as a protein transduction system. This ability appears particularly interesting to induce tissue-specific differentiation when the Tat protein is associated to transcription factors. In the present work, the potential of the complex Tat-MyoD in inducing equine peripheral blood mesenchymal stem cells (PB-MSCs) towards the myogenic fate, was evaluated. Results showed that the internalization process of Tat-MyoD happens only in serum free conditions and that the nuclear localization of the fused complex is observed after 15 hours of incubation. However, the supplement of Tat-MyoD only was not sufficient to induce myogenesis and, therefore, in order to achieve the myogenic differentiation of PB-MSCs, conditioned medium from C2C12 cells was added without direct contact. Real Time PCR and immunofluorescence methods evaluated the establishment of a myogenic program. Our results suggest that TAT- transduction of Tat-MyoD, when supported by conditioned medium, represents a useful methodology to induce myogenic differentiation.
Tat蛋白能够穿过质膜,当它与其他肽融合时,可作为一种蛋白质转导系统。当Tat蛋白与转录因子结合时,这种诱导组织特异性分化的能力显得尤为有趣。在本研究中,评估了Tat-MyoD复合物诱导马外周血间充质干细胞(PB-MSCs)向肌源性命运分化的潜力。结果表明,Tat-MyoD的内化过程仅在无血清条件下发生,且在孵育15小时后观察到融合复合物的核定位。然而,仅添加Tat-MyoD不足以诱导肌生成,因此,为了实现PB-MSCs的肌源性分化,在不直接接触的情况下添加了来自C2C12细胞的条件培养基。实时PCR和免疫荧光方法评估了肌源性程序的建立。我们的结果表明,在条件培养基的支持下,Tat-MyoD的TAT转导是诱导肌源性分化的一种有用方法。