Effect of Rat Bone Marrow Derived-Mesenchymal Stem Cells on Granulocyte Differentiation of Mononuclear Cells as Preclinical Agent in Cellbased Therapy

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. After that, they were cultured, expanded, and characterized. Next, BM-derivedMSCs were co-cultured with MNCs and then were either cultured with 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’s 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.

Granulocyte differentiation of rat bone marrow resident C-kit+ hematopoietic stem cells induced by mesenchymal stem cells could be considered as new option in cell-based therapy

“…In a study by Fathi et al. (2021), it was reported that co-culturing of MNCs with rat BM-derived MSCs cause an increase in the expression levels of granulocyte markers of MNCs, e.g., CD34, CD16, CD11b, and CD18 [ 23 ]. Based on these data, one can conclude that MSCs may affect the granulocyte differentiation of MNCs via ERK protein expression.…”

“…BM mononuclear cells (MNCs) were collected using density gradient centrifugation as previously described by Fathi et al. (2022) [ 23 ]. For this purpose, the ethical code was obtained from the Committee on the Ethics of Tabriz University of Medical Sciences (EthicsNo.…”

In vitro erythrocyte production using human-induced pluripotent stem cells: determining the best hematopoietic stem cell sources

“…Stained samples were measured in a BD Verse flow cytometer (BD Biosciences). FlowJo software version 10.2 (LLC, Ashland, OR, USA) was used to analyze the data [ 26 – 28 ].…”