Isolation of Oct4⁺, Nanog⁺ and SOX2⁻ mesenchymal cells from peripheral blood of a diabetes mellitus patient

Abstract

Diabetes mellitus is a chronic metabolic disorder that affects millions of people worldwide. The most common form is type 2 diabetes mellitus, which results in impaired beta cell function combined with insulin resistance in peripheral organs. One recently proposed treatment approach is the use of adult stem cells derived from bone marrow in autologous stem cell transplantation. Alternatively, peripheral blood can be obtained in a more non-invasive manner. In this study, we isolated and cultured mesenchymal cells (MCs) from the peripheral blood of a diabetes mellitus patient. The cultured cells were large and elongated and had an in vitro migratory capacity in the culture dish. They expressed embryonic stem cell pluripotency markers Nanog and Oct 4 as well as mesenchymal markers CD105 and CD13, and they lacked expression of hematopoietic marker CD45. These characteristics suggest that these cells have a mesenchymal phenotype similar to that obtained from bone marrow cells. The SOX2 gene was downregulated in both the peripheral blood cells and the isolated mesenchymal cell line, indicating a defective mechanism of SOX2 in diabetes mellitus. The overall results of study demonstrate that peripheral blood can be used as a source of MCs from diabetes mellitus patients for use in future regenerative stem cell therapy and that this particular model system may be useful to study the mechanism of diabetes mellitus involving downregulation of the SOX2 cascade.