Vasculogenesis of decidua side population cells of first-trimester pregnancy

Abstract

IntroductionSufficient uterine blood supply is essential for the fetus to develop normally in the uterus. Several mechanisms are involved in the process of vessel development in deciduas and villus. We focus on whether first-trimester decidua side population (SP) cells contain cells capable of differentiating into endothelial cells.MethodsEight decidua samples were collected from healthy women, 22- to 30-years old, undergoing elective terminations of early pregnancy (six to eight gestational weeks). The cell suspensions from human deciduas were stained by Hoechst 33342 and sorted by flow cytometry, further cultured under differentiation conditions and analyzed for specific markers. These cells were implanted into ischemic limbs of nude mice to test the capacity of angiogenesis in vivo by DiI tracers and immunohistochemistry.ResultsDecidua CD31-CD146- SP cells of first-trimester human pregnancy can differentiate into endothelial cells, express the corresponding specific markers of endothelial cells, such as CD31 and CD146, and form tube-like structures on Matrigel and part of newly formed vessels in the ischemic limbs of nude mice. Vascular endothelial growth factor was more effective in promoting proliferation of CD31-CD146-SP cells compared with other growth factors, and estrogen and progesterone at a final concentration of 10 μmol/L and 30 μmol/L, respectively, promoted the migration of CD31-CD146-SP cells in a dose-dependent manner.ConclusionsCD31-CD146- SP cells may be involved in the formation of new vessels in the maternal aspect of the placenta in the first trimester.

Endometrial stem/progenitor cells: the first 10 years

“…The decidua of pregnancy may therefore harbour a subpopulation of undifferentiated MSCs related to eMSCs ( Kyurkchiev et al , 2010 ). Indeed, clonogenic SP cells were identified in the first trimester decidua ( Tsuji et al , 2008 ; Guo et al , 2010 ; Wang et al , 2013 ) comprising 0.03–1.4% of cells, a lower abundance than their endometrial counterparts. SP cells sorted from short-term cultured human decidual cells expressed neither CD31 (endothelial marker) nor CD146 (MSC marker) ( Wang et al , 2013 ), but differentiated into endothelial cells in vitro and induced neovascularization following intramascular injection in a mouse ischaemic hind limb injury model, rescuing the limb ( Wang et al , 2013 ).…”

“…Indeed, clonogenic SP cells were identified in the first trimester decidua ( Tsuji et al , 2008 ; Guo et al , 2010 ; Wang et al , 2013 ) comprising 0.03–1.4% of cells, a lower abundance than their endometrial counterparts. SP cells sorted from short-term cultured human decidual cells expressed neither CD31 (endothelial marker) nor CD146 (MSC marker) ( Wang et al , 2013 ), but differentiated into endothelial cells in vitro and induced neovascularization following intramascular injection in a mouse ischaemic hind limb injury model, rescuing the limb ( Wang et al , 2013 ). This CD31 − CD146 − SP proliferated more rapidly than MP cells when cultured in 0.2% serum-containing media supplemented with either epidermal growth factor (EGF) or fibroblast growth factor 2 (FGF2), similar to clonogenic eMSCs ( Chan et al , 2004 ).…”

Mechanical Isolation of Human Endothelial Cells from Large Vessels for Flow Cytometry Immunophenotyping

“…Here we present an easy and cheap mechanical method to isolate EC from the vessel wall and we tested it by FCM, using a combination of three markers, two of them expressed by EC (CD146/MCAM and CD31/PECAM-1) and one that is not (CD45/LCA [34][35][36][37][38][39]. In a supplementary step, EC identification was reinforced with additional staining with anti-CD309 (VEGFR-2), anti-CD105 (Endoglin) and anti-CD144 (VE-cadherin).…”

“…This method was used to isolate human saphenous vein endothelial cells (HSVEC) in order to identify and characterize them by FCM, using anti-CD45, anti-CD146 and anti-CD31 mAbs and a wash-stain-lyse-no-fix-no-wash procedure. With this staining, we assure the exclusion of other cells that often contaminate the EC suspension and may induce false positive results, such as leukocytes and perivascular cells [34][35][36][37][38][39]. Although nonenzymatic methods have been used for more than 3 decades to isolate EC [9,12,[40][41][42], to the best of our knowledge multi-parametric FCM was never used to characterize the immunophenotype of mechanically isolated fresh EC.…”

The impact of antiandrogen flutamide on the hypoxia inducible factor 1a and vascular endothelial growth factor A gene and protein expression in the pig placenta during late pregnancy

“…The uterine vascular tone has been regulated by progesterone and estrogens. The formation of new blood vessels which occurs in the placenta during early pregnancy is known to be regulated by progesterone [2]. Estrogens, which are locally produced by P450 aromatase within stromal uterine cells, seem to be involved in neovascularization, by increasing the expression of hypoxia inducible factor 2a (HIF-2a) [3].…”