Collagen and Microvascularization in Placentas From Young and Older Mares

In older mares, increasing collagen fibers (fibrosis) in the endometrium and oviduct predisposes to sub-fertility and infertility. In this study, (i) gene transcription of collagen (qPCR: COL1A1, COL1A2, COL3A1, COL5A1); (ii) total collagen protein (hydroxyproline); (iii) collagen distribution (Picrosirius red staining; polarized light microscopy); and (iv) microvascular density (Periodic acid-Schiff staining), were evaluated in mares' placenta, and related to mares age, and placenta and neonate weights. Samples were collected from the gravid horn, non-gravid horn, and body of the placenta from younger (n = 7), and older mares (n = 9) of different breeds. Transcripts of COL1A1, COL3A1 and COL5A1, total collagen protein, chorionic plate connective tissue thickness, and microvascularization increased in the gravid horn of older mares' placentas, compared to the youngest (P < 0.05). Although in other species placenta fibrosis may indicate placental insufficiency and reduced neonate weight, this was not observed here. It appears that older fertile mares, with more parities, may develop a heavier, more vascularized functional placenta with more collagen, throughout a longer gestation, which enables the delivery of heavier foals. Thus, these features might represent morphological and physiological adaptations of older fertile mares' placentas to provide the appropriate nutrition to the equine fetus.

Spatiotemporal Protein Expression Profiles of QSOX1 in the Murine Uterus, Placenta, and Embryo during Pregnancy

Quiescin Q6 sulfhydryl oxidase 1 (QSOX1) catalyzes the oxidation of the sulfhydryl group to disulfide bond and is widely expressed in various tissues. This study focuses on investigating QSOX1′s spatiotemporal and cellular protein expression profile of the pregnant uterus, placenta, and developing embryo during mouse pregnancy. Immunohistochemical staining was used to reveal the localization of QSOX1 protein, and HistoQuest was applied to quantify protein levels. The expression level of QSOX1 in the decidua and muscle cells of the pregnant uterus fluctuated dramatically during pregnancy. QSOX1 was ubiquitously expressed in the labyrinth, junction zone, and chorionic plate in the placenta. The quantitative analysis found that this protein was highly expressed in the spinal cord, lens, midbrain, cerebellum, medulla oblongata, and tooth of mouse embryos, followed by the heart, intercostal muscle, diaphragm, intermediate zone, extrinsic ocular muscle, spine, pons, epidermis, tongue, ganglion, vomeronasal organ, thoracic vertebrae, and thymus. Interestingly, QSOX1 was also markedly expressed in olfactory system tissues. This comprehensive spatiotemporal study of QSOX1 protein expression will provide a basis for further investigations of the QSOX1 physiological function in the pregnant uterus, placenta, and developing embryo.

Placental chorionic plate-derived mesenchymal stem cells ameliorate severe acute pancreatitis by regulating macrophage polarization via secreting TSG-6

Background Mesenchymal stem cells (MSCs) hold promising potential to treat systemic inflammatory diseases including severe acute pancreatitis (SAP). In our previous study, placental chorionic plate-derived MSCs (CP-MSCs) were found to possess superior immunoregulatory capability. However, the therapeutic efficacy of CP-MSCs on SAP and their underlying mechanism remain unclear. Methods The survival and colonization of exogenous CP-MSCs were observed by bioluminescence imaging and CM-Dil labeling in rodent animal models of SAP. The therapeutic efficacy of CP-MSCs on SAP rats was evaluated by pathology scores, the levels of pancreatitis biomarkers as well as the levels of inflammatory factors in the pancreas and serum. The potential protective mechanism of CP-MSCs in SAP rats was explored by selectively depleting M1 or M2 phenotype macrophages and knocking down the expression of TSG-6. Results Exogenous CP-MSCs could survive and colonize in the injured tissue of SAP such as the lung, pancreas, intestine, and liver. Meanwhile, we found that CP-MSCs alleviated pancreatic injury and systemic inflammation by inducing macrophages to polarize from M1 to M2 in SAP rats. Furthermore, our data suggested that CP-MSCs induced M2 polarization of macrophages by secreting TSG-6, and TSG-6 played a vital role in alleviating pancreatic injury and systemic inflammation in SAP rats. Notably, we found that a high inflammation environment could stimulate CP-MSCs to secrete TSG-6. Conclusion Exogenous CP-MSCs tended to colonize in the injured tissue and reduced pancreatic injury and systemic inflammation in SAP rats through inducing M2 polarization of macrophages by secreting TSG-6. Our study provides a new treatment strategy for SAP and initially explains the potential protective mechanism of CP-MSCs on SAP rats.

The Relationship Among Intra-Amniotic Inflammatory Response, The Progression of Inflammation in Chorionic Plate and Early-Onset Neonatal Sepsis

Background: The chorionic plate (CP) has been denigrated by the well-known route of the extraplacental membranes from the decidua parietalis through the chorion to the amnion in the progression of ascending intrauterine infection among preterm births (PTBs). However, considering previous studies reporting the relationship among intra-amniotic inflammatory response (IAIR), the progression of inflammation in extraplacental membranes and early-onset neonatal sepsis (EONS), and the anatomic connection between extraplacental membranes and CP, there is a good chance that IAIR would be more likely and severe according to the progression of inflammation in CP, and this progression of inflammation in CP would be associated with a significant increase in EONS in neonates delivered due to either PTL or preterm-PROM. Unfortunately, there is no information about the relationship among IAIR, the progression of inflammation in CP, and EONS among spontaneous PTBs. The objective of the current study is to examine this issue.Method: The study population included 309 singleton pregnant women-delivered preterm neonates with the following conditions: (1) gestational age (GA) at delivery: 20.0~36.9 weeks; (2) spontaneous PTBs: PTL (151 cases) or preterm-PROM (158 cases); (3) available results of placental histologic examination; (4) without congenital anomaly; and (5) delivery within 60 h of amniocentesis. We examined IAIR, and the frequency of intra-amniotic inflammation (IAI) and EONS according to the progression of inflammation in CP [i.e., stage-0, inflammation-free CP; stage-1, inflammation restricted to subchorionic fibrin (SCF); stage-2, inflammation in connective tissue (CT) of CP but without chorionic vasculitis; and stage-3, chorionic vasculitis]. IAIR was determined by amniotic fluid (AF) matrix metalloproteinase-8 (MMP-8) concentration (ng/ml), and IAI was defined as an elevated AF MMP-8 concentration (≥23 ng/ml). EONS included either suspected or proven EONS.Results: (1) Each stage (stage-0 to stage-3) was present in 69.3% (214/309), 15.9% (49/309), 11.0% (34/309), and 3.9% (12/309) of the study population. (2) AF MMP-8 concentrations continuously elevated according to the progression of inflammation in CP [stage-0 vs. stage-1 vs. stage-2 vs. stage-3; median (ng/ml), range (ng/ml); 6.0 (0.3–4202.7) vs. 153.9 (0.3–6142.6) vs. 464.9 (5.8–3929.0) vs. 1,780.4 (35.1–5019.5); Kruskal–Wallis test, P < 0.001 and Spearman's rank-correlation test, P < 0.000001, r = 0.553]. (3) Moreover, the frequency of IAI and EONS gradually increased with the progression of inflammation in CP [stage-0 vs. stage-1 vs. stage-2 vs. stage-3; IAI, 30.5% (64/210) vs. 70.2% (33/47) vs. 96.7% (29/30) vs. 100% (12/12); EONS, 3.5% (7/200) vs. 25.5% (12/47) vs. 32.3% (10/31) vs. 40.0% (4/10); each for Pearson's chi-square test, P < 0.000001 and linear-by-linear association, P < 0.000001]. (4) Of note, multiple logistic regression analysis demonstrated that a more advanced stage in the progression of inflammation within CP was associated with a higher odds ratio (OR) for EONS [stage-1 vs. stage-2 vs. stage-3; OR, 7.215, 95% confidence-interval (CI) (2.177–23.908) vs. OR, 10.705, 95% CI (2.613–43.849) vs. OR, 27.189, 95% CI (2.557–289.124)] compared with stage-0 even after the adjustment for potential confounding variables.Conclusion: IAIR is more likely and severe according to the progression of inflammation in CP, and this progression of inflammation in CP is an independent risk factor for EONS in spontaneous PTBs. This finding suggests that CP may be another playground for the progression of ascending intrauterine infection in addition to extraplacental membranes, and the progression of inflammation in CP may be used for the prediction of EONS in spontaneous PTBs.

Placental Chorionic Plate-derived Mesenchymal Stem Cells Ameliorate Severe Acute Pancreatitis by Regulating Macrophages Polarization via Secreting TSG-6

BackgroundMesenchymal stem cells (MSCs) hold promising potential to treat systemic inflammatory diseases including severe acute pancreatitis (SAP). In our previous study, placental chorionic plate-derived MSCs (CP-MSCs) were found to possess superior immunoregulatory capability. However, the therapeutic efficacy of CP-MSCs on SAP and their underlying mechanism remain unclear.MethodsThe survival and colonization of exogenous CP-MSCs were observed by bioluminescence imaging and CM-Dil labeling in rodent animal models of SAP. The therapeutic efficacy of CP-MSCs on SAP rats was evaluated by pathology scores, the levels of pancreatitis biomarkers as well as the levels of inflammatory factors in pancreas and serum. The potential protective mechanism of CP-MSCs in SAP rats was explored by selectively depleting M1 or M2 phenotype macrophages and knocking down the expression of TSG-6.ResultsExogenous CP-MSCs could survive and colonize in the injured tissue of SAP such as lung, pancreas, intestine and liver. Meanwhile, we found that CP-MSCs alleviated pancreatic injury and systemic inflammation by inducing macrophages to polarize from M1 to M2 in SAP rats. Furthermore, our data suggested that CP-MSCs induced M2 polarization of macrophages by secreting TSG-6, and TSG-6 played a vital role in alleviating pancreatic injury and systemic inflammation in SAP rats. Notably, we found that a high inflammation environment could stimulate CP-MSCs to secrete TSG-6.ConclusionExogenous CP-MSCs tended to colonize in the injured tissue, and reduced pancreatic injury and systemic inflammation in SAP rats through inducing M2 polarization of macrophages by secreting TSG-6. Our study provides a new treatment strategy for SAP, and initially explains the potential protective mechanism of CP-MSCs on SAP rats.

Isolation and biological characteristics of placental stem cells

Objective. Isolation of stem cells (SCs) from different parts of the placenta and description of their biological characteristics. Materials and methods. The placenta was obtained from parturient women aged 22 to 39 years (mean age was 29.0 ± 3.7 years) with a normal singleton pregnancy resulting in normal delivery and with written informed consent obtained at 37–41 weeks of gestation after non-invasive (n = 32) or operative (n = 2) delivery. Wharton’s jelly from the umbilical cord and the fetal side of the placenta, the chorionic plate with chorion frondosum, were isolated and cultured to obtain mesenchymal stem cells (MSCs). Morphological evaluation of cell culture, differentiation, immunological characterization (CD34, CD45, CD90, CD105, CD73, HLA-DR (BD Biosciences and Becman Coulter, USA)), assessment of viability using 7-ADD dye on the FACSCanto II flow cytometry facility (Becton Dickinson CA, USA), cell growth, sterility control, analysis of STR (Short tandem repeat) loci polymorphism for identification, karyotyping, and cryopreservation of cell cultures were carried out. Results. Primary cell cultures were isolated from the chorionic plate with chorion frondosum and Wharton’s jelly of the umbilical cord in 100% of cases. MSCs had high proliferative activity and viability (98.0 ± 1.2% 7-ADD) throughout the whole period of cultivation. The growth rate of MSCs from the chorionic plate at P3, P4, and P5 was higher compared to that of MSCs from the Wharton’s jelly during cultivation, p ≤ 0.05. Immunophenotype corresponded to the requirements of International Society for Cellular Therapy: high expression of MSC markers (CD73, CD90, CD105) was detected; hematopoietic and lymphocytic markers were absent (CD34, CD45, HLA-DR). The potential of cells to differentiate in the mesenchymal direction (chondrocytes, osteocytes, adipocytes) was confirmed in all obtained MSCs. In 95% of cases, the cells were of fetal origin. The karyotype of the examined MSC cell lines was normal: 46, XX (female) or 46, XY (male). All cell cultures were tested, cryopreserved, and placed in a biobank. Conclusion. Isolation and evaluation of biological characteristics of human placental stem cells is of great interest and is a promising direction in regenerative medicine due to the simplicity of placenta sampling, the absence of ethical problems, the ability to quickly obtain and accumulate the necessary amount of cell material with a stable genotype and given biological characteristics. Key words: mesenchymal stem cells, placenta, immunophenotype, STR loci polymorphisms, karyotype

Vitamin D Promotes Trophoblast Cell Induced Separation of Vascular Smooth Muscle Cells in Vascular Remodeling via Induction of G-CSF

Vitamin D deficiency is associated with complications of pregnancy such as pre-eclampsia, fetal growth restriction, and miscarriage, all of which are also associated with incomplete spiral artery (SpA) remodeling. We have previously shown that both uterine natural killer (uNK) cells and extravillous trophoblast cells (EVT) are required for successful SpA remodeling, but whether their activity in this process is modulated by vitamin D is not known. In the current study, we use a previously described chorionic plate artery (CPA) ex vivo model of vascular remodeling to determine the effects of 1,25(OH)2D treated uNK cell, placental explant (PEx), and uNK/PEx conditioned medium (CM) on vascular smooth muscle cell (VSMC) disorganization and phenotypic switching. Significant results were followed up in VSMCs in vitro. We demonstrate that 1,25(OH)2D can enhance the ability of PEx to induce SpA remodeling, via a mechanism associated with increased secretion of granulocyte-colony stimulating factor (G-CSF). G-CSF appears able to increase VSMC disorganization and phenotypic switching in both an ex vivo vascular model and in vitro VSMC cultures. The clinical relevance of these findings are still to be determined. G-CSF may have differential effects depending on dose and vascular bed, and vitamin D may play a role in potentiating these actions. G-CSF may be an interesting potential therapeutic target for facilitating physiological vascular remodeling for the prevention of adverse obstetric outcomes.