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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) inhibit the proliferation of malignant cells of various hematopoietic origins. (A) The cells (2×104; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured in the presence or absence of the indicated numbers of MSCs in 96-well plates for 48 h. Cell proliferation was assessed using a Counting kit-8 (CCK-8) assay during the final 4 h of culture. (B) The leukemia and lymphoma cells (2×104) were cultured in 96-well plates in the presence of 2×104 MSCs for the indicated periods of time. (C) A20 cells (2×104) were cultured in 96-well plates. MSCs (2×104) were added directly to the A20 cells or on the other side of a Transwell insert, and the plates were co-cultured for 48 h in the presence or absence of an Akt inhibitor (5 µM). A20 cell proliferation was measured using a CCK-8 assay during the final 4 h of culture. The results are shown as a percentage of cell proliferation in comparison with control lymphoma cell proliferation. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance when compared with the control group.
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f1-ijmm-36-01-0139: Mesenchymal stem cells (MSCs) inhibit the proliferation of malignant cells of various hematopoietic origins. (A) The cells (2×104; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured in the presence or absence of the indicated numbers of MSCs in 96-well plates for 48 h. Cell proliferation was assessed using a Counting kit-8 (CCK-8) assay during the final 4 h of culture. (B) The leukemia and lymphoma cells (2×104) were cultured in 96-well plates in the presence of 2×104 MSCs for the indicated periods of time. (C) A20 cells (2×104) were cultured in 96-well plates. MSCs (2×104) were added directly to the A20 cells or on the other side of a Transwell insert, and the plates were co-cultured for 48 h in the presence or absence of an Akt inhibitor (5 µM). A20 cell proliferation was measured using a CCK-8 assay during the final 4 h of culture. The results are shown as a percentage of cell proliferation in comparison with control lymphoma cell proliferation. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance when compared with the control group.

Mentions: We investigated the effects of the MSCs on the prolif-erative activity of leukemia and lymphoma cells of different lineages. The A20 B lymphoma cells (H-2d), the FBL3 erythroleukemia cells (H-2b) and the P388 acute lymphocytic leukemia cells (H-2k) were cultivated with the MSCs for 48 h. When the B6 MSCs were added to the culture in concentrations equivalent to 1 MSC to 1 or 0.4 leukemia cells, the proliferation of the leukemia and lymphoma cells was inhibited; however when the leukemia and lymphoma cells were in excess, the proliferation was not inhibited (Fig. 1A). Furthermore, the anti-proliferative effects of the MSCs increased with increasing co-culture times (Fig. 1B), but these effects were lost when the MSCs and leukemia/lymphoma cells were separated by a permeable membrane (Fig. 1C), indicating that the MSCs inhibited the proliferation of the leukemia and lymphoma cells in a contact-dependent manner. In addition, although it has previously been reported that human MSCs suppress tumor development by inhibiting target-cell Akt activity (6), we did not find that Akt inactivation affected the proliferation of the A20 cells co-cultured with MSCs (Fig. 1C), suggesting that the inhibition of lymphoma cell proliferation by mouse MSCs may not involve the inhibition of Akt.


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) inhibit the proliferation of malignant cells of various hematopoietic origins. (A) The cells (2×104; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured in the presence or absence of the indicated numbers of MSCs in 96-well plates for 48 h. Cell proliferation was assessed using a Counting kit-8 (CCK-8) assay during the final 4 h of culture. (B) The leukemia and lymphoma cells (2×104) were cultured in 96-well plates in the presence of 2×104 MSCs for the indicated periods of time. (C) A20 cells (2×104) were cultured in 96-well plates. MSCs (2×104) were added directly to the A20 cells or on the other side of a Transwell insert, and the plates were co-cultured for 48 h in the presence or absence of an Akt inhibitor (5 µM). A20 cell proliferation was measured using a CCK-8 assay during the final 4 h of culture. The results are shown as a percentage of cell proliferation in comparison with control lymphoma cell proliferation. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance when compared with the control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1-ijmm-36-01-0139: Mesenchymal stem cells (MSCs) inhibit the proliferation of malignant cells of various hematopoietic origins. (A) The cells (2×104; A20 B lymphoma, FBL3 erythroleukemia and P388 acute lymphocytic leukemia cells) were cultured in the presence or absence of the indicated numbers of MSCs in 96-well plates for 48 h. Cell proliferation was assessed using a Counting kit-8 (CCK-8) assay during the final 4 h of culture. (B) The leukemia and lymphoma cells (2×104) were cultured in 96-well plates in the presence of 2×104 MSCs for the indicated periods of time. (C) A20 cells (2×104) were cultured in 96-well plates. MSCs (2×104) were added directly to the A20 cells or on the other side of a Transwell insert, and the plates were co-cultured for 48 h in the presence or absence of an Akt inhibitor (5 µM). A20 cell proliferation was measured using a CCK-8 assay during the final 4 h of culture. The results are shown as a percentage of cell proliferation in comparison with control lymphoma cell proliferation. Results are expressed as the means ± SD of 3 independent experiments. *P<0.05 indicates statistical significance when compared with the control group.
Mentions: We investigated the effects of the MSCs on the prolif-erative activity of leukemia and lymphoma cells of different lineages. The A20 B lymphoma cells (H-2d), the FBL3 erythroleukemia cells (H-2b) and the P388 acute lymphocytic leukemia cells (H-2k) were cultivated with the MSCs for 48 h. When the B6 MSCs were added to the culture in concentrations equivalent to 1 MSC to 1 or 0.4 leukemia cells, the proliferation of the leukemia and lymphoma cells was inhibited; however when the leukemia and lymphoma cells were in excess, the proliferation was not inhibited (Fig. 1A). Furthermore, the anti-proliferative effects of the MSCs increased with increasing co-culture times (Fig. 1B), but these effects were lost when the MSCs and leukemia/lymphoma cells were separated by a permeable membrane (Fig. 1C), indicating that the MSCs inhibited the proliferation of the leukemia and lymphoma cells in a contact-dependent manner. In addition, although it has previously been reported that human MSCs suppress tumor development by inhibiting target-cell Akt activity (6), we did not find that Akt inactivation affected the proliferation of the A20 cells co-cultured with MSCs (Fig. 1C), suggesting that the inhibition of lymphoma cell proliferation by mouse MSCs may not involve the inhibition of Akt.

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