Abstract
Mesenchymal stem cells (MSCs) are widely present in the interstitial tissue of embryos. Although the existence of the metanephros stromal stem cell population has been demonstrated, the focus has been on understanding the process of nephrogenesis, but the biological characterization of stromal stem cell population is less precise. Metanephric mesenchymal stem cells (MMSCs) have a vast potential in kidney tissue engineering and represent excellent candidates in cellular replacement therapy for human disease and veterinary research. Here, we aimed to isolate, culture, and characterize bovine MMSCs. We have successfully obtained a new population of stem cells using renal tissue isolated from three-month-old bovine embryo. MMSCs were isolated by collagenase digestion. The spindle-shaped cells adhered to plastic and exhibited extensive proliferation for more than 26 passages and good clonogenic ability in vitro. Moreover, the metanephric mesenchymal stem cells could be induced to differentiate into mesoderm-derived cells (such as osteoblasts, chondrocytes, and adipocytes) and endoderm-derived hepatocyte-like cells in vitro. These results indicated the multiple differentiations potential of MMSCs. Aside from colony-forming, self-renewal, and multilineage differentiation capabilities, the experiments of immunofluorescence and RT-PCR showed that spindle-shaped cells were positive for MSCs-related markers (CD29, CD44, CD73, CD90, CD106), nestin and vimentin, while the hematopoietic cell surface markers CD34 and pan-leukocyte marker CD45 were undetectable. This study provides a technical platform for the preservation of valuable bovine genetic resources and offers a new source for tissue engineering and cell transplantation therapy.