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Mesenchymal stem cells ameliorate Th1-induced pre-eclampsia-like symptoms in mice via the suppression of TNF-α expression.

Liu L, Zhao G, Fan H, Zhao X, Li P, Wang Z, Hu Y, Hou Y - PLoS ONE (2014)

Bottom Line: The MSCs not only exhibited differentiation and self-renewal capacities, they also possessed immunomodulatory functions and secreted some soluble mediators including IL-6, TGF-β, IDO, VEGF and COX-2.Most importantly, the MSCs were specifically provided with the ability to suppress T cells proliferation by IDO in response to inflammatory cytokine IFN-γ.Strikingly, MSCs-based therapy significantly ameliorated both clinical and histopathological severity of PE symptoms including decreasing the blood pressure and proteinuria, suppressing glomerulonephritis, protecting the feto-placental development.

View Article: PubMed Central - PubMed

Affiliation: Immunology Lab, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.

ABSTRACT
Pre-eclampsia (PE) is thought to be a pregnancy-induced autoimmune disease. Despite several strategies carried out for targeting specific factors relevant to its pathogenesis, PE remains potentially fatal to some patients. Here, we reported a way to isolate mesenchymal stem cells (MSCs) from decidua. The MSCs not only exhibited differentiation and self-renewal capacities, they also possessed immunomodulatory functions and secreted some soluble mediators including IL-6, TGF-β, IDO, VEGF and COX-2. Most importantly, the MSCs were specifically provided with the ability to suppress T cells proliferation by IDO in response to inflammatory cytokine IFN-γ. Moreover, we developed a Th1 cell-induced PE mouse model which displayed a high level of pathogenesis factor TNF-α. Strikingly, MSCs-based therapy significantly ameliorated both clinical and histopathological severity of PE symptoms including decreasing the blood pressure and proteinuria, suppressing glomerulonephritis, protecting the feto-placental development. The therapy also reversed abnormal TNF-α expression in uterine and splenic lymphocytes. These data suggest that MSCs may ameliorate Th1-induced PE-like symptoms in mice via the suppression of TNF-α and MSCs-based therapy may provide a potential novel method for PE.

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Related in: MedlinePlus

Isolation and characterization of MSCs derived from human decidua.(A): Schematic description of the way to isolate MSCs. (B): Morphology of MSCs isolate after 4 day, 6 day and 8 day (20X). (C) Cell growth curve of MSCs. UCMSCs of passage 2 were plated in 24-well plates in DMEM/F12 supplemented with 10% FBS at a density of 5×103 cells/well. The cells were detached with 0.25% trypsin-EDTA every 2 days and cell counting until 14 day. Bars show the mean. n = 3. (D) Osteogenic differentiation was assayed by the alizarin red staining. No mineralized matrix formation was found in UCMSCs maintained in regular growth medium. Osteogenic differentiation was evidenced by the formation of mineralized matrix in UCMSCs after osteogeneic induction. (10X). Scale bars, 100 µm. (E) Adipogenic differentiation was detected by oil red O staining. No lipid vacuoles were found in in UCMSCs maintained in the regular medium. Adipogenic differentiation was evidenced by the formation of lipid vacuoles by oil-red O staining in UCMSCs after adipogenic induction. (10X) Scale bars, 100 µm. All data are representative of three independent experiments.
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pone-0088036-g001: Isolation and characterization of MSCs derived from human decidua.(A): Schematic description of the way to isolate MSCs. (B): Morphology of MSCs isolate after 4 day, 6 day and 8 day (20X). (C) Cell growth curve of MSCs. UCMSCs of passage 2 were plated in 24-well plates in DMEM/F12 supplemented with 10% FBS at a density of 5×103 cells/well. The cells were detached with 0.25% trypsin-EDTA every 2 days and cell counting until 14 day. Bars show the mean. n = 3. (D) Osteogenic differentiation was assayed by the alizarin red staining. No mineralized matrix formation was found in UCMSCs maintained in regular growth medium. Osteogenic differentiation was evidenced by the formation of mineralized matrix in UCMSCs after osteogeneic induction. (10X). Scale bars, 100 µm. (E) Adipogenic differentiation was detected by oil red O staining. No lipid vacuoles were found in in UCMSCs maintained in the regular medium. Adipogenic differentiation was evidenced by the formation of lipid vacuoles by oil-red O staining in UCMSCs after adipogenic induction. (10X) Scale bars, 100 µm. All data are representative of three independent experiments.

Mentions: A novel way was developed for isolating MSCs from human decidua here.In general, an enzyme cocktail (hyaluronidase 5 U/ml, collagenase 125 U/ml and dispase 50 U/ml; Sigma) instead of collagenase was used here to digest the fragments of decidua. After washing twice, large pieces of tissue in the digested mixture did not need to be removed and the digested mixture also did not need to pass through a 100 µm filter to obtain single cell suspensions [Fig. 1A]. Adherent cells with fibroblastic morphology could be observed as early as 24 hours. The cells formed a monolayer of homogenous bipolar spindle-like cells with a whirl pool like array within 1 week [Fig. 1B]. Then, MSCs were serially passaged to examine their expansion potential and determined to expand readily for up to 2 weeks [Fig. 1C]. After 3 cell passages, the adherent cells were symmetric with phenotypic surface antigens, which is positive for CD105, CD73, CD90, HLA-ABC, CD29, CD44 and negative for HLA-DR, CD19, CD11b, CD14, CD34, CD31 (date not show). In addition, our obtained MSCs possessed the abilities of osteogenic differentiation [Fig. 1D] and adipogenic differentiation [Fig. 1E], which detected by the calcification of the matrix (alizarin red staining) and oil red O staining, respectively.


Mesenchymal stem cells ameliorate Th1-induced pre-eclampsia-like symptoms in mice via the suppression of TNF-α expression.

Liu L, Zhao G, Fan H, Zhao X, Li P, Wang Z, Hu Y, Hou Y - PLoS ONE (2014)

Isolation and characterization of MSCs derived from human decidua.(A): Schematic description of the way to isolate MSCs. (B): Morphology of MSCs isolate after 4 day, 6 day and 8 day (20X). (C) Cell growth curve of MSCs. UCMSCs of passage 2 were plated in 24-well plates in DMEM/F12 supplemented with 10% FBS at a density of 5×103 cells/well. The cells were detached with 0.25% trypsin-EDTA every 2 days and cell counting until 14 day. Bars show the mean. n = 3. (D) Osteogenic differentiation was assayed by the alizarin red staining. No mineralized matrix formation was found in UCMSCs maintained in regular growth medium. Osteogenic differentiation was evidenced by the formation of mineralized matrix in UCMSCs after osteogeneic induction. (10X). Scale bars, 100 µm. (E) Adipogenic differentiation was detected by oil red O staining. No lipid vacuoles were found in in UCMSCs maintained in the regular medium. Adipogenic differentiation was evidenced by the formation of lipid vacuoles by oil-red O staining in UCMSCs after adipogenic induction. (10X) Scale bars, 100 µm. All data are representative of three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928118&req=5

pone-0088036-g001: Isolation and characterization of MSCs derived from human decidua.(A): Schematic description of the way to isolate MSCs. (B): Morphology of MSCs isolate after 4 day, 6 day and 8 day (20X). (C) Cell growth curve of MSCs. UCMSCs of passage 2 were plated in 24-well plates in DMEM/F12 supplemented with 10% FBS at a density of 5×103 cells/well. The cells were detached with 0.25% trypsin-EDTA every 2 days and cell counting until 14 day. Bars show the mean. n = 3. (D) Osteogenic differentiation was assayed by the alizarin red staining. No mineralized matrix formation was found in UCMSCs maintained in regular growth medium. Osteogenic differentiation was evidenced by the formation of mineralized matrix in UCMSCs after osteogeneic induction. (10X). Scale bars, 100 µm. (E) Adipogenic differentiation was detected by oil red O staining. No lipid vacuoles were found in in UCMSCs maintained in the regular medium. Adipogenic differentiation was evidenced by the formation of lipid vacuoles by oil-red O staining in UCMSCs after adipogenic induction. (10X) Scale bars, 100 µm. All data are representative of three independent experiments.
Mentions: A novel way was developed for isolating MSCs from human decidua here.In general, an enzyme cocktail (hyaluronidase 5 U/ml, collagenase 125 U/ml and dispase 50 U/ml; Sigma) instead of collagenase was used here to digest the fragments of decidua. After washing twice, large pieces of tissue in the digested mixture did not need to be removed and the digested mixture also did not need to pass through a 100 µm filter to obtain single cell suspensions [Fig. 1A]. Adherent cells with fibroblastic morphology could be observed as early as 24 hours. The cells formed a monolayer of homogenous bipolar spindle-like cells with a whirl pool like array within 1 week [Fig. 1B]. Then, MSCs were serially passaged to examine their expansion potential and determined to expand readily for up to 2 weeks [Fig. 1C]. After 3 cell passages, the adherent cells were symmetric with phenotypic surface antigens, which is positive for CD105, CD73, CD90, HLA-ABC, CD29, CD44 and negative for HLA-DR, CD19, CD11b, CD14, CD34, CD31 (date not show). In addition, our obtained MSCs possessed the abilities of osteogenic differentiation [Fig. 1D] and adipogenic differentiation [Fig. 1E], which detected by the calcification of the matrix (alizarin red staining) and oil red O staining, respectively.

Bottom Line: The MSCs not only exhibited differentiation and self-renewal capacities, they also possessed immunomodulatory functions and secreted some soluble mediators including IL-6, TGF-β, IDO, VEGF and COX-2.Most importantly, the MSCs were specifically provided with the ability to suppress T cells proliferation by IDO in response to inflammatory cytokine IFN-γ.Strikingly, MSCs-based therapy significantly ameliorated both clinical and histopathological severity of PE symptoms including decreasing the blood pressure and proteinuria, suppressing glomerulonephritis, protecting the feto-placental development.

View Article: PubMed Central - PubMed

Affiliation: Immunology Lab, Medical School and State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing, China.

ABSTRACT
Pre-eclampsia (PE) is thought to be a pregnancy-induced autoimmune disease. Despite several strategies carried out for targeting specific factors relevant to its pathogenesis, PE remains potentially fatal to some patients. Here, we reported a way to isolate mesenchymal stem cells (MSCs) from decidua. The MSCs not only exhibited differentiation and self-renewal capacities, they also possessed immunomodulatory functions and secreted some soluble mediators including IL-6, TGF-β, IDO, VEGF and COX-2. Most importantly, the MSCs were specifically provided with the ability to suppress T cells proliferation by IDO in response to inflammatory cytokine IFN-γ. Moreover, we developed a Th1 cell-induced PE mouse model which displayed a high level of pathogenesis factor TNF-α. Strikingly, MSCs-based therapy significantly ameliorated both clinical and histopathological severity of PE symptoms including decreasing the blood pressure and proteinuria, suppressing glomerulonephritis, protecting the feto-placental development. The therapy also reversed abnormal TNF-α expression in uterine and splenic lymphocytes. These data suggest that MSCs may ameliorate Th1-induced PE-like symptoms in mice via the suppression of TNF-α and MSCs-based therapy may provide a potential novel method for PE.

Show MeSH
Related in: MedlinePlus