Conditioned medium obtained from placental mesenchymal stem cells and use thereof in the therapeutic treatment of preeclampsia

Abstract

It is described a conditioned medium (CM) obtainable by culturing, in a liquid culture medium, placental mesenchymal stem cells isolated from the placental tissue of pregnant women not affected by preeclampsia. The conditioned medium of the invention includes at least IL-6, IL-10 and MCP-1 proteins and it is effective for the therapeutic treatment of preeclampsia.

1 . A conditioned medium obtainable by in vitro culturing an isolated placental mesenchymal stem cell population from the placenta of a pregnant woman not affected by preeclampsia in a basal serum-free liquid culture medium for at least three hours, the conditioned medium comprising at least the cell-secreted factors interleukin-6 (IL-6), interleukin-10 (IL-10) and monocyte chemoattractant protein 1 (MCP-1). 
     
     
         2 . The conditioned medium according to  claim 1 , comprising a further cell-secreted factor selected from the group consisting of ENA-78, GCSF, GRO, GRO-alpha, IL-6, IL-7, IL-8, MCP-1, MCP-2, MCSF, MDC, ANGIOGENIN, ONCOSTATIN m, VEGF, BDNF, BLC, CKb 8-1, EOTAXIN 2, EOTAXIN 3, FLT-3 LIGAND, FRACTALKINE, GCP-2, GDNF, HGF, IGFBP-1, IGFBP-2, IGFBP-4, IP-10, LIF, LIGHT, MCP-4, MIF, MIP-3alpha, NAP-2, NT-3, OSTEOPONTIN, OSTOPROTEGERIN, TGF-beta 2, TIMP-1, TIMP-2, RANTES, IGFBP-3, IL1b, IL-3, MIP-1b, PIGF, IL-1a, I309, FGF9, TARC, PDGF-bb, LEPTIN, TNF-α and any combination thereof. 
     
     
         3 . The conditioned medium according to  claim 2 , wherein the further cell-secreted factor is interleukin 8 (IL-8). 
     
     
         4 . The conditioned medium according to any of  claim 1 , wherein the placental stem cell has a chorionic or amniotic origin. 
     
     
         5 . The conditioned medium according to any of  claim 1 , which is cell free. 
     
     
         6 . A method of preparing a conditioned medium according to  claim 1 , comprising the steps of:
 (i) in vitro culturing an isolated placental mesenchymal stem cell population from the placenta of a pregnant woman not affected by preeclampsia in a serum-free liquid basal culture medium for at least 3 hours;   ii) separating the cell fraction from the liquid culture medium, thereby obtaining a conditioned medium comprising a combination of factors secreted by said placental mesenchymal stem cells, said combination comprising at least interleukin-6 (IL-6), interleukin-10 (IL-10) and monocyte chemoattractant protein 1 (MCP-1).   
     
     
         7 . The method according to  claim 6 , wherein said conditioned medium comprises a further cell-secreted factor selected from the group consisting of ENA-78, GCSF, GRO, GRO-alpha, IL-6, IL-7, IL-8, MCP-1, MCP-2, MCSF, MDC, ANGIOGENIN, ONCOSTATIN m, VEGF, BDNF, BLC, CKb 8-1, EOTAXIN 2, EOTAXIN 3, FLT-3 LIGAND, FRACTALKINE, GCP-2, GDNF, HGF, IGFBP-1, IGFBP-2, IGFBP-4, IP-10, LIF, LIGHT, MCP-4, MIF, MIP-3alpha, NAP-2, NT-3, OSTEOPONTIN, OSTOPROTEGERIN, TGF-beta 2, TIMP-1, TIMP-2, RANTES, IGFBP-3, IL1b, IL-3, MIP-1b, PIGF, IL-1a, I309, FGF9, TARC, PDGF-bb, LEPTIN, TNF-α and any combination thereof. 
     
     
         8 . The method according  claim 6 , wherein the further cell-secreted factor is IL-8. 
     
     
         9 . The method according to  claim 6 , wherein the culturing time of the placental mesenchymal stem cells from the placenta of a pregnant woman not affected by preeclampsia is of at least 6 hours, or at least 12 hours, or at least 48 hours, or at least 72 hours. 
     
     
         10 . The method according to  claim 6 , wherein the cell fraction is separated from the liquid culture medium by filtration and/or centrifugation. 
     
     
         11 . The method according to  claim 6 , further comprising step iv) of isolating from the conditioned medium obtained in step iii) one or more of the factors secreted by said placental mesenchymal stem cells. 
     
     
         12 . The method according to  claim 11 , wherein step iv) comprises isolating the IL-6, IL-10, MCP-1, ENA-78, GCSF, GRO, GRO-alpha, IL-6, IL-7, IL-8, MCP-1, MCP-2, MCSF, MDC, ANGIOGENIN, ONCOSTATIN m, VEGF, BDNF, BLC, CKb 8-1, EOTAXIN 2, EOTAXIN 3, FLT-3 LIGAND, FRACTALKINE, GCP-2, GDNF, HGF, IGFBP-1, IGFBP-2, IGFBP-4, IP-10, LIF, LIGHT, MCP-4, MIF, MIP-3alpha, NAP-2, NT-3, OSTEOPONTIN, OSTOPROTEGERIN, TGF-beta 2, TIMP-1, TIMP-2, RANTES, IGFBP-3, IL1b, IL-3, MIP-1b, PIGF, IL-1a, I309, FGF9, TARC, PDGF-bb, LEPTIN, TNF-α factor or any combination thereof. 
     
     
         13 . A, method of therapeutically treating preeclampsia comprising administering the conditioned medium according to  claim 1  to a subject in need thereof. 
     
     
         14 . The method according to  claim 13 , the conditioned medium is in a pharmaceutical form suitable for systemic administration. 
     
     
         15 . The method according to  claim 14 , wherein the pharmaceutical form is suitable for administration by injection. 
     
     
         16 . A method of antinflammatory treatment, comprising administering to a subject in need thereof a medicament comprising the conditioned medium according to  claim 1 . 
     
     
         17 . The method according to  claim 16 , wherein the subject is affected by preeclampsia.