RESUMO
Objective: umbilical cord blood is used for transplantation purposes in regenerative medicine of hematological disorders. MicroRNAs are important regulators of gene expression that control both physiological and pathological processes such as cancer development and incidence. There is a new relation between p53 [tumor suppressor gene] and miR-145 [suppressor of cell growth] upregulation. In this study, we have assessed how adipose-derived stem cells [ADSCs] affect the expansion of hematopoietic stem cells [HSCs], as well as miR-145 and p53 expressions
Materials and Methods: in this experimental study, we cultured passage-3 isolated human ADSCs as a feeder layer. Flow cytometry analysis confirmed the presence of ADSC surface markers CD73, CD90, CD105. Ex vivo cultures of cordblood CD34+ cells were cultured under the following 4 culture conditions for 7 days: i. Medium only supplemented with cytokines, ii. Culture on an ADSCs feeder layer, iii. Indirect culture on an ADSCs feeder layer [Thin Cert[] plate with a 0.4 ?m pore size], and iv. Control group analyzed immediately after extraction. Real-time polymerase chain reaction [PCR] was used to determine the expressions of the p53 and miR-145 genes. Flow cytometry analysis of cells stained by annexin V and propidium iodide [PI] was performed to detect the rate of apoptosis in the expanded cells
Results: ADSCs tested positive for mesenchymal stem cell [MSC] markers CD105, CD90, and CD73, and negative for HSC markers CD34 and CD45. Our data demonstrated the differentiation potential of ASCs to osteoblasts by alizarin red and alkaline phosphatase staining. MTT assay results showed a higher proliferation rate of CD34+cells directly cultured on the ADSCs feeder layer group compared to the other groups. Direct contact between HSCs and the feeder layer was prevented by a microporous membrane p53 expression increased in the HSCs group with indirect contact of the feeder layer compared to direct contact of the feeder layer. p53 significantly downregulated in HSCs cultured on ADSCs, whereas miR-145 significantly upregulated in HSCs cultured on ADSCs
Conclusion: ADSCs might increase HSCs proliferation and self-renewal through miR-145, p53, and their relationship
RESUMO
Type 1 diabetes mellitus [T1DM] is a disease where destruction of the insulin producing pancreatic beta-cells leads to increased blood sugar levels. Both genetic and environmental factors play a part in the development of T1DM. Currently, numerous loci are specified to be the responsible genetic factors for T1DM; however, the mechanisms of only a few of these genes are known. Although several environmental factors are presumed responsible for progression of T1DM, to date, most of their mechanisms remain undiscovered. After several years of hyperglycemia, late onsets of macrovascular [e.g., cardiovascular] and microvascular [e.g., neurological, ophthalmological, and renal] complications may occur. This review and accompanying figures provides an overview of the etiological factors for T1DM, its pathogenesis at the cellular level, and attributed complications