Effect of Rat Bone Marrow Derived-Mesenchymal Stem Cells on Granulocyte Differentiation of Mononuclear Cells as Preclinical Agent in Cellbased Therapy
Ezzatollah Fathi, Sheyda Azarbad, Raheleh Farahzadi, Sara Javanmardi, Ilja Vietor
Abstract
BACKGROUND: Bone marrow mononuclear cells (BM-MNCs), as a collection of hematopoietic and mesenchymal stem cells (MSCs), are capable of producing all blood cell lineages. The use of cytokines, growth factors or cells capable of secreting these factors will help in stimulating the proliferation and differentiation of these cells into mature cell lines. On the other hand, MSCs are multipotent stromal cells that can be differentiated into various cell lineages. Moreover, these cells can control the process of hematopoiesis by secreting cytokines and growth factors. The present study aimed to investigate the effect of BM-derived MSCs on the differentiation of MNCs based on the assessment of cell surface markers by flow cytometry analysis. METHODS: For this purpose, the MNCs were purified from rat BM using density gradient centrifugation. Thereafter, they were cultured, expanded, and characterized. Next, BM-derived-MSCs were cocultured with MNCs, and then were either cultured MNCs alone (control group) or co-cultured MNCs with BM-derived-MSCs (experimental group). Finally, they were collected on day 7 and subjected to flow cytometry analysis for granulocyte markers and ERK protein investigation. RESULTS: It was found that the expression levels of CD34, CD16, CD11b, and CD18 granulocyte markers as well as protein expression of ERK have significantly increased in the experimental group compared to the control group. CONCLUSION: Therefore, it can be concluded that MSCs could affect the granulocyte differentiation of MNCs via ERK protein expression, which is a key component of the ERK signaling pathway.