Improvement of Paracrine potentialities of Mesenchymal stem cells using simple suspension culturing method

Introduction: Mesenchymal stem cells (MSCs) are isolated from different tissues including (WJ). They have shown to have paracrine ability to support hematopoietic stem cells (HSCs) expansion in vitro. However, the WJ-derived MSCs are very heterogeneous and have limited capacity to secret cytokines that involve in hematopoiesis. Hence, improvement of their culture condition might promote the secretion capacity of WJ-MSCs. Materials and methods: Herein, we set up a simple culturing method for WJ-MSCs which promotes their cytokines secretion. WJ-MSCs were separated from wharton's jelly. The isolated cells were added to Methocult medium diluted in α-MEM and seeded in poly-HEMA-coated plates. WJ-MSCs conditioned medium was harvested and concentrated from serum free media. The expression of some cytokines including IL-6, IL-11, granulocyte-macrophage colony-stimulating factor (GM-CSF) was evaluated by RT-PCR and western blot techniques. WJ-MSCs that cultivated under suspension condition expressed more GM-CSF and IL-11 in comparison to those of that cultured in adherent conditions (P<0.05 and P<0.001). Results: There was no significant difference between the secreted amount of IL-6 from suspended and adherent WJ-MSCs. Suspension cultivation of WJ-MSCs enhances their ability to produce some important hematopoietic cytokines. Conclusion: These suspended WJ-MSCs could be used for more effective low cost in vitro expansion of HSCs

کلیدواژه ها:

Wharton's jelly ، MSCs ، Paracrine effects ، HSCs engraftment