Contributions
Type: Publication Only
Background
Mesenchymal Stem Cells (MSCs) are a group of adult stem cells naturally found in the body. These cells were reported from many sources. Bone marrow and adipose tissue are two main sources of human MSCs. MSCs have attracted considerable attention in the fields of cell and Gene Therapy due to their intrinsic characteristics and ability to differentiate into multiple lineages. Whereas BM has been the first recognized source of MSC, adipose tissue represents a valid reservoir of mesenchymal progenitors. Adipose tissue can be obtained in relevant amount and easily processed to release large numbers of adipose-derived MSC (AD-MSC). These cells may offer efficient tools for cell-based Gene Therapy approaches.
Aims
In this study, we evaluated whether AD-MSC could deliver TRAIL for leukemic cells treatment and induce apoptosis in leukemic cells.
Methods
Human AD-MSCs were isolated, characterized and transduced with a lentiviral vector encoding secretory tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). TRAIL protein production verified by western blotting. TRAIL-ADMSCs were Co-cultured with HL-60 and AML-M3 patients MNCs. Apoptosis induction was studied by flow cytometry using annexin-FITC. We also studied MSCs migration potential by Trans-Well method.
Results
In coculture experiment of TRAIL secreting MSCs with leukemic cells we observed TRAIL-AD-MSC targeted HL-60 cells and AML-M3 mononuclear cells (MNCs). We observed significant apoptosis inducing potential by TRAIL-ADMSC in HL-60 cells and AML patients' mononuclear cells (MNCs) compared to control once (P < 0.05). Maximum apoptosis induction in HL-60 cells was 25% which shows most of these cells have resistance to TRAIL induced cell death. Whatever in patients MNCs maximum induction of apoptosis was 45%. In vitro coculture experiments on Trans-well plate's migration assays showed that HL-60 cells culture but not FBS lacked growth media, supported the migration of hMSCs and enhanced their migration (P < 0.05).
Summary
We found that stably TRAIL transduced ADMSCs could serve as constant source of TRAIL production. However this treatment modality didn’t completely induced apoptosis in all HL-60 or MNCs nevertheless it indicate needs for finding cotreatments appropriate to type of resistance mechanism. We also observed that leukemic cells are able to production and releasing cytokines which cause migration of AD-MSCs toward leukemic cells and also AD-MSCs have relevant receptors on the cell surface to respond such cytokines.
In conclusion these results suggest that human AD-MSCs have potential use as effective delivery vehicles for therapeutic genes in the treatment of hematologic malignancies.
Keyword(s): Apoptosis, Mesenchymal stem cell, Migration, TRAIL
Session topic: Publication Only
Type: Publication Only
Background
Mesenchymal Stem Cells (MSCs) are a group of adult stem cells naturally found in the body. These cells were reported from many sources. Bone marrow and adipose tissue are two main sources of human MSCs. MSCs have attracted considerable attention in the fields of cell and Gene Therapy due to their intrinsic characteristics and ability to differentiate into multiple lineages. Whereas BM has been the first recognized source of MSC, adipose tissue represents a valid reservoir of mesenchymal progenitors. Adipose tissue can be obtained in relevant amount and easily processed to release large numbers of adipose-derived MSC (AD-MSC). These cells may offer efficient tools for cell-based Gene Therapy approaches.
Aims
In this study, we evaluated whether AD-MSC could deliver TRAIL for leukemic cells treatment and induce apoptosis in leukemic cells.
Methods
Human AD-MSCs were isolated, characterized and transduced with a lentiviral vector encoding secretory tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). TRAIL protein production verified by western blotting. TRAIL-ADMSCs were Co-cultured with HL-60 and AML-M3 patients MNCs. Apoptosis induction was studied by flow cytometry using annexin-FITC. We also studied MSCs migration potential by Trans-Well method.
Results
In coculture experiment of TRAIL secreting MSCs with leukemic cells we observed TRAIL-AD-MSC targeted HL-60 cells and AML-M3 mononuclear cells (MNCs). We observed significant apoptosis inducing potential by TRAIL-ADMSC in HL-60 cells and AML patients' mononuclear cells (MNCs) compared to control once (P < 0.05). Maximum apoptosis induction in HL-60 cells was 25% which shows most of these cells have resistance to TRAIL induced cell death. Whatever in patients MNCs maximum induction of apoptosis was 45%. In vitro coculture experiments on Trans-well plate's migration assays showed that HL-60 cells culture but not FBS lacked growth media, supported the migration of hMSCs and enhanced their migration (P < 0.05).
Summary
We found that stably TRAIL transduced ADMSCs could serve as constant source of TRAIL production. However this treatment modality didn’t completely induced apoptosis in all HL-60 or MNCs nevertheless it indicate needs for finding cotreatments appropriate to type of resistance mechanism. We also observed that leukemic cells are able to production and releasing cytokines which cause migration of AD-MSCs toward leukemic cells and also AD-MSCs have relevant receptors on the cell surface to respond such cytokines.
In conclusion these results suggest that human AD-MSCs have potential use as effective delivery vehicles for therapeutic genes in the treatment of hematologic malignancies.
Keyword(s): Apoptosis, Mesenchymal stem cell, Migration, TRAIL
Session topic: Publication Only