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Mouse bone marrow-derived mesenchymal stem cells inhibit leukemia/lymphoma cell proliferation in vitro and in a mouse model of allogeneic bone marrow transplant.

Song N, Gao L, Qiu H, Huang C, Cheng H, Zhou H, Lv S, Chen L, Wang J - Int. J. Mol. Med. (2015)

Bottom Line: Cell proliferation, apoptosis, cell cycle progression and the amount of cytokine secretion were then measured using a Cell Counting kit‑8, Annexin V/propidium iodide staining, flow cytometry and ELISA, respectively.Our results revealed that the MSCs inhibited the proliferation of the mouse lymphoma and leukemia cells in vitro, leading to cell cycle arrest and reducing the secretion of interleukin (IL)‑10.The results from the present study indicate that MSCs may be safe and effective when used in allogeneic BMT for the treatment of hemotological malignancies.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Institute of Hematology, PLA, Changhai Hospital, Second Military Medical University, Shanghai 200433, P.R. China.

ABSTRACT
The allogeneic hematopoietic stem cell (HSC) transplantation of mesenchymal stem cells (MSCs) contributes to the reconstitution of hematopoiesis by ameliorating acute graft‑versus‑host disease (aGVHD). However, the role of MSCs in graft‑versus‑leukemia remains to be determined. In the present study, we co‑cultured C57BL/6 mouse bone marrow (BM)‑derived MSCs with A20 murine B lymphoma, FBL3 murine erythroleukemia and P388 murine acute lymphocytic leukemia cells. Cell proliferation, apoptosis, cell cycle progression and the amount of cytokine secretion were then measured using a Cell Counting kit‑8, Annexin V/propidium iodide staining, flow cytometry and ELISA, respectively. We also established a model of allogeneic bone marrow transplantation (BMT) using BALB/c mice. Following the administration of A20 cells and MSCs, we recorded the symptoms and the survival of the mice for 4 weeks, assessed the T cell subsets present in peripheral blood, and, after the mice were sacrifice, we determined the infiltration of MSCs into the organs by histological staining. Our results revealed that the MSCs inhibited the proliferation of the mouse lymphoma and leukemia cells in vitro, leading to cell cycle arrest and reducing the secretion of interleukin (IL)‑10. In our model of allogeneic BMT, the intravenous injection of MSCs into the mice injected wth A20 cells decreased the incidence of lymphoma, improved survival, increased the fraction of CD3+CD8+ T cells, decreased the fraction of CD3+CD4+ T cells and CD4+CD25+ T cells in peripheral blood, and ameliorated the manifestation of aGVHD. The results from the present study indicate that MSCs may be safe and effective when used in allogeneic BMT for the treatment of hemotological malignancies.

No MeSH data available.


Related in: MedlinePlus

Mesenchymal stem cells (MSCs) induce the early apoptosis of leukemia and lymphoma cells. (A) Cells (1×105; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured for 72 h alone or in the presence of MSCs at ratio of 1 MSC to 2 leukemia/lymphoma cells. The cells were then harvested and analyzed by Annexin V and propidium iodide (PI) double staining. (B) A20 cells (1×105) were cultured under 3 different conditions in 6-well plates for 72 h: with or without MSCs (ratio 1:1, respectively); some were physically separated from teh MSCs using a Transwell system. (C) p21 and caspase-3 mRNA expression in A20, FBL3, and P388 cells detected by RT-qPCR. p21 and caspase-3 mRNA expression was increased in the cells in co-culture relative to culture alone. The data are expressed as the means ± SD of 3 separate experiments. *P<0.05 indicates statistical significance.
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f2-ijmm-36-01-0139: Mesenchymal stem cells (MSCs) induce the early apoptosis of leukemia and lymphoma cells. (A) Cells (1×105; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured for 72 h alone or in the presence of MSCs at ratio of 1 MSC to 2 leukemia/lymphoma cells. The cells were then harvested and analyzed by Annexin V and propidium iodide (PI) double staining. (B) A20 cells (1×105) were cultured under 3 different conditions in 6-well plates for 72 h: with or without MSCs (ratio 1:1, respectively); some were physically separated from teh MSCs using a Transwell system. (C) p21 and caspase-3 mRNA expression in A20, FBL3, and P388 cells detected by RT-qPCR. p21 and caspase-3 mRNA expression was increased in the cells in co-culture relative to culture alone. The data are expressed as the means ± SD of 3 separate experiments. *P<0.05 indicates statistical significance.

Mentions: When the A20 B lymphoma cells, the FBL3 erythroleukemia cells and the P388 acute lymphocytic leukemia cells were co-cultured with the MSCs for 72 h, the proportion of apoptotic cells, as measured by Annexin V and PI staining, was significantly increased (Fig. 2A and B). The proportion of cells in the G0/G1 phase was also significantly higher in the cells incubated with the MSCs than in those incubated alone, while the proportion of cells in the S phase was significantly decreased following co-culture with the MSCs for 72 h (Fig. 3A and Table I). When the MSCs were physically separated from the A20 cells using a Transwell system, they no longer influenced the early apoptotic rate of the A20 cells (Fig. 2B) or the cell cycle (Fig. 3B), suggesting that MSCs influence the cell cycle and apoptosis of lymphoma cells in a contact-dependent manner.


Mouse bone marrow-derived mesenchymal stem cells inhibit leukemia/lymphoma cell proliferation in vitro and in a mouse model of allogeneic bone marrow transplant.

Song N, Gao L, Qiu H, Huang C, Cheng H, Zhou H, Lv S, Chen L, Wang J - Int. J. Mol. Med. (2015)

Mesenchymal stem cells (MSCs) induce the early apoptosis of leukemia and lymphoma cells. (A) Cells (1×105; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured for 72 h alone or in the presence of MSCs at ratio of 1 MSC to 2 leukemia/lymphoma cells. The cells were then harvested and analyzed by Annexin V and propidium iodide (PI) double staining. (B) A20 cells (1×105) were cultured under 3 different conditions in 6-well plates for 72 h: with or without MSCs (ratio 1:1, respectively); some were physically separated from teh MSCs using a Transwell system. (C) p21 and caspase-3 mRNA expression in A20, FBL3, and P388 cells detected by RT-qPCR. p21 and caspase-3 mRNA expression was increased in the cells in co-culture relative to culture alone. The data are expressed as the means ± SD of 3 separate experiments. *P<0.05 indicates statistical significance.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494598&req=5

f2-ijmm-36-01-0139: Mesenchymal stem cells (MSCs) induce the early apoptosis of leukemia and lymphoma cells. (A) Cells (1×105; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured for 72 h alone or in the presence of MSCs at ratio of 1 MSC to 2 leukemia/lymphoma cells. The cells were then harvested and analyzed by Annexin V and propidium iodide (PI) double staining. (B) A20 cells (1×105) were cultured under 3 different conditions in 6-well plates for 72 h: with or without MSCs (ratio 1:1, respectively); some were physically separated from teh MSCs using a Transwell system. (C) p21 and caspase-3 mRNA expression in A20, FBL3, and P388 cells detected by RT-qPCR. p21 and caspase-3 mRNA expression was increased in the cells in co-culture relative to culture alone. The data are expressed as the means ± SD of 3 separate experiments. *P<0.05 indicates statistical significance.
Mentions: When the A20 B lymphoma cells, the FBL3 erythroleukemia cells and the P388 acute lymphocytic leukemia cells were co-cultured with the MSCs for 72 h, the proportion of apoptotic cells, as measured by Annexin V and PI staining, was significantly increased (Fig. 2A and B). The proportion of cells in the G0/G1 phase was also significantly higher in the cells incubated with the MSCs than in those incubated alone, while the proportion of cells in the S phase was significantly decreased following co-culture with the MSCs for 72 h (Fig. 3A and Table I). When the MSCs were physically separated from the A20 cells using a Transwell system, they no longer influenced the early apoptotic rate of the A20 cells (Fig. 2B) or the cell cycle (Fig. 3B), suggesting that MSCs influence the cell cycle and apoptosis of lymphoma cells in a contact-dependent manner.

Bottom Line: Cell proliferation, apoptosis, cell cycle progression and the amount of cytokine secretion were then measured using a Cell Counting kit‑8, Annexin V/propidium iodide staining, flow cytometry and ELISA, respectively.Our results revealed that the MSCs inhibited the proliferation of the mouse lymphoma and leukemia cells in vitro, leading to cell cycle arrest and reducing the secretion of interleukin (IL)‑10.The results from the present study indicate that MSCs may be safe and effective when used in allogeneic BMT for the treatment of hemotological malignancies.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Institute of Hematology, PLA, Changhai Hospital, Second Military Medical University, Shanghai 200433, P.R. China.

ABSTRACT
The allogeneic hematopoietic stem cell (HSC) transplantation of mesenchymal stem cells (MSCs) contributes to the reconstitution of hematopoiesis by ameliorating acute graft‑versus‑host disease (aGVHD). However, the role of MSCs in graft‑versus‑leukemia remains to be determined. In the present study, we co‑cultured C57BL/6 mouse bone marrow (BM)‑derived MSCs with A20 murine B lymphoma, FBL3 murine erythroleukemia and P388 murine acute lymphocytic leukemia cells. Cell proliferation, apoptosis, cell cycle progression and the amount of cytokine secretion were then measured using a Cell Counting kit‑8, Annexin V/propidium iodide staining, flow cytometry and ELISA, respectively. We also established a model of allogeneic bone marrow transplantation (BMT) using BALB/c mice. Following the administration of A20 cells and MSCs, we recorded the symptoms and the survival of the mice for 4 weeks, assessed the T cell subsets present in peripheral blood, and, after the mice were sacrifice, we determined the infiltration of MSCs into the organs by histological staining. Our results revealed that the MSCs inhibited the proliferation of the mouse lymphoma and leukemia cells in vitro, leading to cell cycle arrest and reducing the secretion of interleukin (IL)‑10. In our model of allogeneic BMT, the intravenous injection of MSCs into the mice injected wth A20 cells decreased the incidence of lymphoma, improved survival, increased the fraction of CD3+CD8+ T cells, decreased the fraction of CD3+CD4+ T cells and CD4+CD25+ T cells in peripheral blood, and ameliorated the manifestation of aGVHD. The results from the present study indicate that MSCs may be safe and effective when used in allogeneic BMT for the treatment of hemotological malignancies.

No MeSH data available.


Related in: MedlinePlus