Cytokines secreted from bone marrow derived mesenchymal stem cells promote apoptosis and change cell cycle distribution of K562 cell line as clinical agent in cell transplantation

Abstract

Mesenchymal stem cells (MSCs) are of special interest due their potential clinical use in cell-based therapy. Therapies engaging MSCs are showing increasing promise in the cancer treatment and anticancer drug screening applications. A multitude of growth factors and cytokines secreted from these cells are known to give such multifunctional properties, but details of their role are yet to be absolutely demonstrated. In this study, we have evaluated the influence of BMSCs on K562 cell line as chronic myeloid leukemia (CML) cells, with the use of a cytokine antibody array recognizing 34 cytokines. For this purpose, BMSCs were isolated and co-cultured with K562 cells; thereafter, cultured K562 alone and co-cultured K562 with BMSCs (10:1) were collected at day 7 and subjected to cell cycle distribution assay as well as annexin/PI analysis and Ki/caspase-3 assay for apoptosis assessment. In the following, the gene and protein expression levels of BAX and BCL-2 as pro- and anti-apoptotic agents were investigated. Furthermore, after 7 days’ treatment, culture medium was collected from both control and experimental groups for cytokine antibody array. It was found that BMSCs resulted in a robust increase in the number of cells at G 0 /G 1 phase and arrest the G 0 /G 1 phase as well as significantly inducing late apoptosis in K562 cells. The significant presence of TIMP-1 (tissue inhibitor of metalloproteinases-1), and moderate elevated signals for CINC-1 (cytokine-induced neutrophil chemoattractant-1) were obvious in the co-cultured conditioned media, but no significant increase was found in 32 other cytokines. It is concluded that co-culture of BMSCs with K562 cells could secrete a substantial amount of TIMP-1 and CINC-1. These cytokines could be involved in the inhibition of the K562 cell proliferation via BAX and caspase-3 cascade pathways.

Human Dental Pulp Stem Cells Exhibit Osteogenic Differentiation Potential

“…We proliferative potential and ability to differentiate into osteoblasts and therefore could be used for regenerative medicine applications in oral and maxillofacial surgery. CFU-F as described by Gronthos et al [22] Huang et al [23] and El-Gendy et al [1]. The high proliferative potential of DPSCs indicate their potential use in applications such as tissue engineering and regenerative medicine.…”

“…selected explant culture method as it is inexpensive and gives a pure population of stem cells [21]. Within 3-4 days after primary culture, a certain number of cells (DPSCs) appeared in culture that resembled MSCs as defined by their spindle shape, fibroblastic morphology and plasticadherent growth as documented previously [22,23]. The International Society for Cellular Therapy (ISCT) defines the criteria for characterization of MSCs.…”

Cardiac differentiation of bone marrow resident c-kit+ stem cells by L-carnitine increases through secretion of VEGF, IL6, IGF-1 and TGF-β

“…In a preliminary study by Freed et al (2005), it was shown that upregulation of cardiotrophin-1 (CT-1) as a member of the IL-6 cytokine family inhibited the fibroblast proliferation in the infarcted myocardium zone [39]. In previous study by Tang [44]. The output data were processed with FlowJo software version X.0.7.…”

<p>Muscone Promotes The Adipogenic Differentiation Of Human Gingival Mesenchymal Stem Cells By Inhibiting The Wnt/β-Catenin Signaling Pathway</p>

“…Flow cytometry was used to determine the fluorescence intensity and number of positive cells. To determine the growth rate of GMSCs in the exponential phase, the population doubling time (PDT) was calculated: PDT = CT/PDN, where CT is the culture time, and PDN = log (N1/N0) ×3.31 (N1, final cell number; N0, initial cell number) 28. To examine the multidirectional differentiation potential of GMSCs,29,30 GMSCs were cultured in osteogenesis-inducing medium (α-MEM supplemented with 10% FBS, 1 μM dexamethasone, 50 mg/L ascorbic acid, and 3 mM β-glycerophosphate) (Sigma-Aldrich, St. Louis, MO, USA) or adipogenesis-inducing medium (α-MEM supplemented with 10% FBS, 0.5 mM IBMX, 1 μM dexamethasone, 200 μM indomethacin, and 10 μM insulin) (Sigma-Aldrich, St. Louis, MO, USA).…”