Elsevier

Placenta

Volume 36, Issue 3, March 2015, Pages 312-321
Placenta

Mesenchymal stem cells reside in a vascular niche in the decidua basalis and are absent in remodelled spiral arterioles

https://doi.org/10.1016/j.placenta.2014.12.014Get rights and content

Highlights

  • DMSCs were isolated from the decidua basalis region attached to the placenta.

  • The maternal origin of DMSCs was verified.

  • DMSCs expressed MSC markers and differentiated into mesodermal lineages.

  • DMSCs were localised to the vascular niche of non-transformed spiral arterioles of the decidua.

  • In fully transformed spiral arterioles, DMSCs were absent from the vascular niche.

Abstract

Introduction

Maternal decidua basalis tissue attached to the placenta following delivery is a source of decidual mesenchymal stem cells (DMSCs). The in vitro characteristics of DMSCs have been partly defined but their in vivo function(s) are poorly understood. The anatomic location, or niche, provides clues regarding potential in vivo function(s) of DMSCs, but the niche has not been described.

Methods

Cells were isolated from the decidua basalis and flow cytometric analyses showed the expected phenotypic profile for MSC cell surface markers. In vitro, the cells differentiated into adipocytes, osteocytes, and chondrocytes. DMSCs were then stained with antibodies by immunofluorescence detection.

Results

Immunocytochemistry revealed that DMSCs were positive for FZD-9, STRO-1, 3G5, and α-SMA as expected and lacked expression of vWF and Ck7. Fluorescence in situ hybridization analysis showed the cultured cells were of maternal origin. Immunofluorescence was carried out on placental bed biopsies using the FZD-9, STRO-1, 3G5, and α-SMA antibodies. DMSCs were located in the vascular niche in decidua basalis. Immunofluorescence with antibodies to FZD-9, Ck7 and vWF revealed DMSCs in the vascular niche surrounding intact non-transformed spiral arterioles but DMSCs were absent in fully transformed spiral arterioles.

Discussion

Spiral arteriole remodelling is a critical feature of human pregnancy. The DMSC niche was investigated in fully transformed and non-transformed spiral arterioles. DMSCs have not been previously implicated in spiral arteriole remodelling. The absence of DMSCs around fully transformed spiral arterioles suggests they are a target for replacement or destruction by invading placental extravillous trophoblast cells, which carry out spiral arteriole remodelling.

Introduction

The decidua basalis is a thin plate of maternal endometrial tissue that is structurally and functionally transformed by pregnancy and forms the attachment site of the placenta. During delivery, the decidua basalis adhering to the placenta is torn away from the underlying maternal tissues and remains attached to the placenta.

The decidua basalis and human placenta are rich sources of maternal and fetal derived mesenchymal stem cells (MSCs), respectively [1], [2], [3]. These cells are also referred to as mesenchymal stromal cells. Unlike placental MSCs, which are well characterised in vitro and are already in preclinical studies and clinical trials [4], [5], [6], [7], the in vitro properties and potential clinical applications of maternal decidua basalis MSCs (DMSCs) are less well known. Most important to this work is that our understanding of the in vivo function(s) of DMSCs is rudimentary. Identifying the stem cell niche can provide important clues as to the in vivo function(s) of DMSCs.

The stem cell niche is defined as the specialised local microenvironment where stem cells reside and the niche contributes directly toward stem cell maintenance [8]. Identifying the MSC niche in tissues is fundamental to understanding MSC function but locating the niche in human tissues is challenging. The retention of bromodeoxyuridine label by slow-cycling MSCs has been widely utilized to locate these cells in vivo [8], [9], [10]. MSC niches in bone marrow, periodontal ligament, dental pulp, and human adipose tissue were identified by employing a combination of characteristic cell surface markers [11], [12], [13], [14], [15], [16]. Combinations of markers are essential since no single, specific marker for MSCs has been identified. This probably reflects the heterogeneity of MSC populations in vivo, which adds further complexity to assigning the niche [17]. Some markers used to identify MSCs in a vascular niche are expressed in smooth muscle cells (α-smooth muscle actin/α-SMA), and pericytes (CD146, STRO-1, 3G5, frizzled-9/FZD-9). Using a combination of markers (CD146, VLA-1/CD49a, STRO-1, and 3G5), we localised fetal MSCs to a vascular niche within the chorionic villi of the placenta [18], consistent with vascular niches described for MSCs in many other organs and tissues [17].

The decidua basalis plays a critical role in human pregnancy, but the niche for MSCs in this tissue has not yet been identified. Early in human pregnancy (8–10 weeks), specialised invasive fetal extravillous trophoblast cells (EVTs) detach from the tips of fetal chorionic villi, invade into the decidua basalis and underlying myometrium, and migrate along the spiral arteries. EVTs infiltrate and replace, completely or partially, the vascular smooth muscle cells surrounding the spiral arterioles, and the endothelial cells that line the lumen of the spiral arterioles. This process remodels the spiral arterioles into large diameter conduit vessels of low resistance, which facilitates increased blood flow to the intervillous space of the placenta allowing it to meet the increasing nutritional demands of the rapidly growing fetus. Thus, remodelling of maternal spiral arterioles is essential for successful pregnancy [19], [20]. Identification of the DMSC niche may provide important clues as to the role DMSCs play in placental development, and specifically in spiral arteriole remodelling.

In this study, we employed a strategy similar to our previous studies on placental MSC niches [18], [21]. First, DMSCs were isolated from the decidua basalis according to published methods [22], [23], [24]. Next, the cultured cells were assessed for MSC criteria proposed by the International Society of Cellular Therapy (ISCT) [25]. These criteria include that MSCs must adhere to plastic surfaces, should express CD105, CD73 and CD90 but not CD34, CD14, CD19, CD11b, CD79α or HLA-DR, and have multipotent differentiation potential [25]. Further criteria set by Parolini et al. for placental MSCs include the determination of the fetal or maternal origin of the cells [26]. Then, immunocytochemistry was performed on cultured DMSCs to determine whether they were positive for markers used to define MSC niches (FZD-9, 3G5, α-SMA, and STRO-1). Finally, multi-label immunofluorescence staining of placental bed biopsies with this combination of niche markers was used to identify the DMSC niche in the decidua basalis before and after spiral arteriole remodelling.

Section snippets

Tissue collection

Placental samples were collected from healthy women with medically uncomplicated pregnancies following Caesarean section or vaginal delivery at term (n = 5). The tissue had the typical morphology of a healthy placenta with no obvious signs of calcification, infarcts or meconium staining. Exclusion criteria for tissue collection were twin or triplet pregnancy, maternal smoking, drug dependency, intrauterine infection, prolonged rupture of the fetal membranes, and placental abruption. Informed

Isolation and characterization of DMSCs

Cells were isolated from the decidua basalis attached to villous placental tissue collected from women with uncomplicated pregnancies delivered by Caesarean section or vaginal delivery at term. The cells adhered to tissue culture flasks and had the fibroblast-like appearance typical of MSCs (Fig. 1A). The yield of DMSCs at P0 was not determined because the cell population was not homogeneous. At P1, when the population displayed a homogeneous, fibroblast-like morphology, the yield was 5 × 105

Discussion

Cells were successfully isolated from the decidua basalis that remains attached to the placenta following delivery, using a tissue mincing and enzymatic digestion method. Following in vitro growth and passaging, the cells appeared homogeneous and exhibited the characteristic fibroblastic morphology of MSCs (Fig. 1Ai). Cells prepared met accepted ISCT criteria for MSCs, including adherence to plastic cultureware, surface marker expression, and differentiation potential [25].

Immunophenotyping

Conflict of interest

The author(s) declared that there is no potential conflict of interest with respect to the research, authorship, and/or publication of this article.

Acknowledgements

We acknowledge the patients who consented to provide their samples, the obstetric staff who provided the placental bed biopsy samples, and the clinical research midwives at the Royal Women's Hospital, Sue Duggan and Moira Stewart for sample collection. We thank Melissa Duggan, Dr. Maria Kokkinos, and May Grgurinovic for their excellent technical assistance. This work was supported by grant funds from King Abdullah International Medical Research Centre (Grant No. RC08/114), the Royal Women's

References (52)

  • M. Dominici et al.

    Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement

    Cytotherapy

    (2006)
  • A.C. Staff et al.

    Augmented PLA2Activity in pre-eclamptic decidual tissue—a key player in the pathophysiology of ‘Acute Atherosis’ in pre-eclampsia?

    Placenta

    (2003)
  • S.D. Smith et al.

    Evidence for immune cell involvement in decidual spiral arteriole remodeling in early human pregnancy

    Am J Pathol

    (2009)
  • C.B. Portmann-Lanz et al.

    Placental mesenchymal stem cells as potential autologous graft for pre- and perinatal neuroregeneration

    Am J Obstet Gynecol

    (2006)
  • X. Zhang et al.

    Successful immortalization of mesenchymal progenitor cells derived from human placenta and the differentiation abilities of immortalized cells

    Biochem Biophys Res Commun

    (2006)
  • V.L. Battula et al.

    Human placenta and bone marrow derived MSC cultured in serum-free, b-FGF-containing medium express cell surface frizzled-9 and SSEA-4 and give rise to multilineage differentiation

    Differentiation

    (2007)
  • H. Ning et al.

    Mesenchymal stem cell marker Stro-1 is a 75 kd endothelial antigen

    Biochem Biophys Res Commun

    (2011)
  • F. Lyall

    Priming and remodelling of human placental bed spiral arteries during pregnancy–a review

    Placenta

    (2005)
  • Y. Khong et al.

    Defective deep placentation

    Best Pract Res Clin Obstet Gynaecol

    (2011)
  • R. Ohlsson et al.

    PDGFB regulates the development of the labyrinthine layer of the mouse fetal placenta

    Dev Biol

    (1999)
  • I. Brosens et al.

    The great obstetrical syndromes are associated with disorders of deep placentation

    Am J Obstet Gynecol

    (2011)
  • C.W.G. Redman

    Preeclampsia: a multi-stress disorder

    Rev Méd Interne

    (2011)
  • J.H. Hwang et al.

    Cytokine expression in placenta-derived mesenchymal stem cells in patients with pre-eclampsia and normal pregnancies

    Cytokine

    (2010)
  • Y. Fukuchi et al.

    Human placenta-derived cells have mesenchymal stem/progenitor cell potential

    Stem Cells

    (2004)
  • M.H. Abumaree et al.

    Phenotypic and functional characterization of mesenchymal stem cells from chorionic villi of human term placenta

    Stem Cell Rev Rep

    (2013)
  • M.H. Abumaree et al.

    Human placental mesenchymal stem cells (pMSCs) play a role as immune suppressive cells by shifting macrophage differentiation from inflammatory M1 to anti-inflammatory M2 macrophages

    Stem Cell Rev

    (2013)
  • Cited by (34)

    • Decidual mesenchymal stem/stromal cells from preeclamptic patients secrete endoglin, which at high levels inhibits endothelial cell attachment in vitro

      2022, Placenta
      Citation Excerpt :

      In the vascular niche, DMSCs share opposite sides of a common basement membrane with endothelial cells. Furthermore, we showed that DMSCs are also replaced or destroyed by EVTs in spiral arteriole remodelling [5,6]. Aberrant placental development is a hallmark of PE [7].

    • The fate of human SUSD2+ endometrial mesenchymal stem cells during decidualization

      2022, Stem Cell Research
      Citation Excerpt :

      It has been previously reported that eMSC remain quiescent in the perivascular niche (Schwab et al., 2005, Schwab and Gargett, 2007, Masuda et al., 2012). In decidua, Kusuma et al (2015) located perivascular FZD-9 positive MSC only in the non-transformed arterioles, suggesting that during arteriolar transformation, perivascular MSC are destroyed (Kusuma et al., 2015). Our analysis revealed a statistically significant lower abundance of SUSD2+ eMSC in the 1st trim and P compared to the NP group.

    • Decidual mesenchymal stem/stromal cell-derived extracellular vesicles ameliorate endothelial cell proliferation, inflammation, and oxidative stress in a cell culture model of preeclampsia

      2020, Pregnancy Hypertension
      Citation Excerpt :

      The MSC phenotype was confirmed by flow cytometry using MSC cell surface marker antibodies (Table 2). MSC cell surface markers were as previously published [15]. Flow cytometry was performed on an LSR Fortessa X-20 cytometer and data were analyzed using Weasel software v3.3.3.

    View all citing articles on Scopus
    View full text