Limits...
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

Co-culture of A20 B lymphoma cells and mesenchymal stem cells (MSCs) causes a decrease in the levels of interleukin (IL)-10 in the supernatant. A20 cells (1×105) were cultured in 6-well plates in the presence or absence of MSCs at ratio 1:1 for 24, 48, or 72 h. (A) IL-10 levels in A20 culture supernatants were measured by ELISA in the presence or absence of MSCs for 72 h. (B) The fraction of CD19-positive A20 cells expressing intracellular IL-10 was measured by flow cytometry. (C) The levels of transforming growth factor (TGF)-β, tumor necrosis factor (TNF)-α and interferon (IFN)-γ in the culture supernatants were measured by ELISA. (D) The fraction of cells containing intracellular IL-10 was measured by flow cytometry. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4494598&req=5

f4-ijmm-36-01-0139: Co-culture of A20 B lymphoma cells and mesenchymal stem cells (MSCs) causes a decrease in the levels of interleukin (IL)-10 in the supernatant. A20 cells (1×105) were cultured in 6-well plates in the presence or absence of MSCs at ratio 1:1 for 24, 48, or 72 h. (A) IL-10 levels in A20 culture supernatants were measured by ELISA in the presence or absence of MSCs for 72 h. (B) The fraction of CD19-positive A20 cells expressing intracellular IL-10 was measured by flow cytometry. (C) The levels of transforming growth factor (TGF)-β, tumor necrosis factor (TNF)-α and interferon (IFN)-γ in the culture supernatants were measured by ELISA. (D) The fraction of cells containing intracellular IL-10 was measured by flow cytometry. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance.

Mentions: As cytokines play an important role in the regulation of adaptive and innate immune responses to tumors (12–14), we measured the contents of cytokines in the A20 cells incubated in the presence or absence of MSCs. The A20 cells were found to express low levels of TNF-α and IFN-γ, moderate levels of TGF-β, and high levels of IL-10 (Fig. 4A and B). When co-cultured with the MSCs at a ratio of 1:1, the levels of IL-10 in the supernatant were significantly decreased, and the inhibitory effects of the MSCs on IL-10 secretion became more prominent with time (Fig. 4A). However, the levels of TGF-β, TNF-α, and IFN-γ in the supernatant were not affected by the presence of MSCs (Fig. 4C).


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)

Co-culture of A20 B lymphoma cells and mesenchymal stem cells (MSCs) causes a decrease in the levels of interleukin (IL)-10 in the supernatant. A20 cells (1×105) were cultured in 6-well plates in the presence or absence of MSCs at ratio 1:1 for 24, 48, or 72 h. (A) IL-10 levels in A20 culture supernatants were measured by ELISA in the presence or absence of MSCs for 72 h. (B) The fraction of CD19-positive A20 cells expressing intracellular IL-10 was measured by flow cytometry. (C) The levels of transforming growth factor (TGF)-β, tumor necrosis factor (TNF)-α and interferon (IFN)-γ in the culture supernatants were measured by ELISA. (D) The fraction of cells containing intracellular IL-10 was measured by flow cytometry. Results are expressed as the means ± SD of 3 independent 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

f4-ijmm-36-01-0139: Co-culture of A20 B lymphoma cells and mesenchymal stem cells (MSCs) causes a decrease in the levels of interleukin (IL)-10 in the supernatant. A20 cells (1×105) were cultured in 6-well plates in the presence or absence of MSCs at ratio 1:1 for 24, 48, or 72 h. (A) IL-10 levels in A20 culture supernatants were measured by ELISA in the presence or absence of MSCs for 72 h. (B) The fraction of CD19-positive A20 cells expressing intracellular IL-10 was measured by flow cytometry. (C) The levels of transforming growth factor (TGF)-β, tumor necrosis factor (TNF)-α and interferon (IFN)-γ in the culture supernatants were measured by ELISA. (D) The fraction of cells containing intracellular IL-10 was measured by flow cytometry. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance.
Mentions: As cytokines play an important role in the regulation of adaptive and innate immune responses to tumors (12–14), we measured the contents of cytokines in the A20 cells incubated in the presence or absence of MSCs. The A20 cells were found to express low levels of TNF-α and IFN-γ, moderate levels of TGF-β, and high levels of IL-10 (Fig. 4A and B). When co-cultured with the MSCs at a ratio of 1:1, the levels of IL-10 in the supernatant were significantly decreased, and the inhibitory effects of the MSCs on IL-10 secretion became more prominent with time (Fig. 4A). However, the levels of TGF-β, TNF-α, and IFN-γ in the supernatant were not affected by the presence of MSCs (Fig. 4C).

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