Fathi Ezzatollah, Azarbad Sheyda, Farahzadi Raheleh, Javanmardi Sara, Vietor Ilja
Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Curr Gene Ther. 2022;22(2):152-161. doi: 10.2174/1566523221666210519111933.
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.
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.
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.
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.
骨髓单个核细胞(BM - MNCs)作为造血干细胞和间充质干细胞(MSCs)的集合,能够产生所有血细胞谱系。使用细胞因子、生长因子或能够分泌这些因子的细胞将有助于刺激这些细胞增殖并分化为成熟细胞系。另一方面,间充质干细胞是多能基质细胞,可分化为各种细胞谱系。此外,这些细胞可通过分泌细胞因子和生长因子来控制造血过程。本研究旨在通过流式细胞术分析评估细胞表面标志物,来研究骨髓来源的间充质干细胞对单个核细胞分化的影响。
为此,使用密度梯度离心法从大鼠骨髓中纯化单个核细胞。此后,对其进行培养、扩增和鉴定。接下来,将骨髓来源的间充质干细胞与单个核细胞共培养,然后将单个核细胞单独培养(对照组)或将单个核细胞与骨髓来源的间充质干细胞共培养(实验组)。最后,在第7天收集细胞,并进行流式细胞术分析以检测粒细胞标志物和ERK蛋白。
发现与对照组相比,实验组中CD34、CD16、CD11b和CD18粒细胞标志物的表达水平以及ERK的蛋白表达均显著增加。
因此,可以得出结论,间充质干细胞可通过ERK蛋白表达影响单个核细胞的粒细胞分化,而ERK蛋白是ERK信号通路的关键组成部分。