Human chorionic mesenchymal stem/stromal cells (CMSCs) derived from the placenta are similar to adult tissue-derived MSCs. The aim of this study was to investigate the role of these cells in normal placental development. Transcription factors, particularly members of the homeobox gene family, play crucial roles in maintaining stem cell proliferation and lineage specification in embryonic tissues. In adult tissues and organs, stem cells proliferate at low levels in their niche until they receive cues from the microenvironment to differentiate. The homeobox genes that are expressed in the CMSC niche in placental tissues have not been identified. We used the novel strategy of laser capture microdissection to isolate the stromal component of first trimester villi and excluded the cytotrophoblast and syncytiotrophoblast layers that comprise the outer layer of the chorionic villi. Microarray analysis was then used to screen for homeobox genes in the microdissected tissue. Candidate homeobox genes were selected for further RNA analysis. Immunohistochemistry of candidate genes in first trimester placental villous stromal tissue revealed homeobox genes Meis1, myeloid ectropic viral integration site 1 homolog 2 (MEIS2), H2.0-like Drosophila (HLX), transforming growth factor β-induced factor (TGIF), and distal-less homeobox 5 (DLX5) were expressed in the vascular niche where CMSCs have been shown to reside. Expression of MEIS2, HLX, TGIF, and DLX5 was also detected in scattered stromal cells. Real-time polymerase chain reaction and immunocytochemistry verified expression of MEIS2, HLX, TGIF, and DLX5 homeobox genes in first trimester and term CMSCs. These data suggest a combination of regulatory homeobox genes is expressed in CMSCs from early placental development to term, which may be required for stem cell proliferation and differentiation.
LongoUGLoppiniMBertonALa VerdeLKhanWSDenaroV. Stem cells from umbilical cord and placenta for musculoskeletal tissue engineering. Curr Stem Cell Res Ther. 2012;7(4):272–281.
3.
ManuelpillaiUMoodleyYBorlonganCVParoliniO. Amniotic membrane and amniotic cells: potential therapeutic tools to combat tissue inflammation and fibrosis?Placenta. 2011;32(suppl 4):S320–S325.
4.
ParoliniOAlvianoFBagnaraGP. Concise review: isolation and characterization of cells from human term placenta: outcome of the first international workshop on placenta derived stem cells. Stem Cells. 2008;26(2):300–311.
5.
In ’t AnkerPSScherjonSAKleijburg-van der KeurC. Isolation of mesenchymal stem cells of fetal or maternal origin from human placenta. Stem Cells. 2004;22(7):1338–1345.
6.
AbumareeMHAl JumahMAKalionisB. Phenotypic and functional characterization of mesenchymal stem cells from chorionic villi of human term placenta. Stem Cell Rev. 2013;9(1):6.
7.
CastrechiniNMMurthiPGudeNM. Mesenchymal stem cells in human placental chorionic villi reside in a vascular Niche. Placenta. 2010;31(3):203–212.
8.
JonesGNMoschidouDPuga-IglesiasTI. Ontological differences in first compared to third trimester human fetal placental chorionic stem cells. PLoS One. 2012;7(9):e43395.
9.
ArakawaRAokiRArakawaMSaitoK. Human first-trimester chorionic villi have a myogenic potential. Cell Tissue Res. 2012;348(1):189–197.
10.
FukuchiYNakajimaHSugiyamaDHiroseIKitamuraTTsujiK. Human placenta-derived cells have mesenchymal stem/progenitor cell potential. Stem Cells. 2004;22(5):649–658.
11.
MaltepeEBakardjievAIFisherSJ. The placenta: transcriptional, epigenetic, and physiological integration during development. J Clin Invest. 2010;120(4):1016–1025.
12.
CrossJC. How to make a placenta: mechanisms of trophoblast cell differentiation in mice--a review. Placenta. 2005;26(suppl A):S3–S9.
13.
CrossJCBaczykDDobricN. Genes, development and evolution of the placenta. Placenta. 2003;24(2-3):123–130.
PanGThomsonJA. Nanog and transcriptional networks in embryonic stem cell pluripotency. Cell Res. 2007;17(1):42–49.
16.
LoreggerTPollheimerJKnoflerM. Regulatory transcription factors controlling function and differentiation of human trophoblast--a review. Placenta. 2003;24(suppl A):S104–S110.
17.
KuboHShimizuMTayaY. Identification of mesenchymal stem cell (MSC)-transcription factors by microarray and knockdown analyses, and signature molecule-marked MSC in bone marrow by immunohistochemistry. Genes Cells. 2009;14(3):407–424.
18.
KlumpHSchiedlmeierBBaumC. Control of self-renewal and differentiation of hematopoietic stem cells: HOXB4 on the threshold. Ann N Y Acad Sci. 2005;1044:6–15.
19.
TroskoJE.From adult stem cells to cancer stem cells: Oct-4 Gene, cell-cell communication, and hormones during tumor promotion. Ann N Y Acad Sci. 2006;1089:36–58.
20.
TolkunovaECavaleriFEckardtS. The caudal-related protein cdx2 promotes trophoblast differentiation of mouse embryonic stem cells. Stem Cells. 2006;24(1):139–144.
21.
NiwaHToyookaYShimosatoD. Interaction between Oct3/4 and Cdx2 determines trophectoderm differentiation. Cell. 2005;123(5):917–929.
22.
ChenLYabuuchiAEminliS. Cross-regulation of the Nanog and Cdx2 promoters. Cell Res. 2009;19(9):1052–1061.
23.
AckemaKBChariteJ. Mesenchymal stem cells from different organs are characterized by distinct topographic Hox codes. Stem Cells Dev. 2008;17(5):979–991.
NikolovaGStrilicBLammertE. The vascular niche and its basement membrane. Trends Cell Biol. 2007;17(1):19–25.
26.
CastrechiniNMMurthiPQinS. Decidua parietalis-derived mesenchymal stromal cells reside in a vascular niche within the choriodecidua. Reprod Sci. 2012;19(12):1302–1314.
27.
MartiniMMJeremias TdaSKohlerMCMarosticaLLTrentinAGAlvarez-SilvaM. Human placenta-derived mesenchymal stem cells acquire neural phenotype under the appropriate niche conditions. DNA Cell Biol. 2013;32(2):58–65.
ShahdadfarAFronsdalKHaugTReinholtFPBrinchmannJE. In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability. Stem Cells. 2005;23(9):1357–1366.
30.
Nur FarihaMMChuaKHTanGCTanAEHayatiAR. Human chorion-derived stem cells: changes in stem cell properties during serial passage. Cytotherapy. 2011;13(5):582–593.
31.
BilbanMTauberSHaslingerP. Trophoblast invasion: assessment of cellular models using gene expression signatures. Placenta. 2010;31(11):989–996.
32.
PoloniARosiniVMondiniE. Characterization and expansion of mesenchymal progenitor cells from first-trimester chorionic villi of human placenta. Cytotherapy. 2008;10(7):690–697.
33.
HawesCSPetropoulosALopataAKalionisBJonesWR. Reactivity of human trophoblast monoclonal antibodies with marmoset monkey trophoblast cultures. Hum Reprod. 1998;13(5):1169–1174.
34.
LivakKJSchmittgenTD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402–408.
35.
HollandPWBoothHABrufordEA. Classification and nomenclature of all human homeobox genes. BMC Biol. 2007;5:47.
36.
BattulaVLTremlSAbeleHBuhringHJ. Prospective isolation and characterization of mesenchymal stem cells from human placenta using a frizzled-9-specific monoclonal antibody. Differentiation. 2008;76(4):326–336.
37.
RiderDANalathambyTNurcombeVCoolSM. Selection using the alpha-1 integrin (CD49a) enhances the multipotentiality of the mesenchymal stem cell population from heterogeneous bone marrow stromal cells. J Mol Histol. 2007;38(5):449–458.
38.
LiuGClementLCKanwarYSAvila-CasadoCChughSS. ZHX proteins regulate podocyte gene expression during the development of nephrotic syndrome. J Biol Chem. 2006;281(51):39681–39692.
39.
BuhringHJBattulaVLTremlS. Novel markers for the prospective isolation of human MSC. Ann N Y Acad Sci. 2007;1106:262–271.
40.
da Silva MeirellesLChagastellesPCNardiNB. Mesenchymal stem cells reside in virtually all post-natal organs and tissues. J Cell Sci. 2006;119(pt 11):2204–2213.
41.
RajaramanGMurthiPQuinnLBrenneckeSPKalionisB. Homeodomain protein HLX is expressed primarily in cytotrophoblast cell types in the early pregnancy human placenta. Reprod Fertil Dev. 2008;20(3):357–367.
42.
PathirageNACocquebertMSadovskyY. Homeobox gene transforming growth factor beta-induced factor-1 (TGIF-1) is a regulator of villous trophoblast differentiation and its expression is increased in human idiopathic fetal growth restriction. Mol Hum Reprod. 2013;19(10):665–675.
43.
MurthiPSoMGudeNMDohertyVLBrenneckeSPKalionisB. Homeobox genes are differentially expressed in macrovascular human umbilical vein endothelial cells and microvascular placental endothelial cells. Placenta. 2007;28(2-3):219–223.
44.
MurthiPHidenURajaramanG. Novel homeobox genes are differentially expressed in placental microvascular endothelial cells compared with macrovascular cells. Placenta. 2008;29(7):624–630.
45.
SungHJHongSCYooJH. Stemness evaluation of mesenchymal stem cells from placentas according to developmental stage: comparison to those from adult bone marrow. J Korean Med Sci. 2010;25(10):1418–1426.
46.
Portmann-LanzCBBaumannMUMuellerM. Neurogenic characteristics of placental stem cells in preeclampsia. Am J Obstet Gynecol. 2010;203(4):399.e391–397.
47.
RajaramanGMurthiPPathirageNBrenneckeSPKalionisB. Downstream targets of homeobox gene HLX show altered expression in human idiopathic fetal growth restriction. Am J Pathol. 2010;176(1):278–287.
48.
RajaramanGMurthiPLeoBBrenneckeSPKalionisB. Homeobox gene HLX1 is a regulator of colony stimulating factor-1 dependent trophoblast cell proliferation. Placenta. 2007;28(10):991–998.
49.
PaigeSLThomasSStoick-CooperCL. A temporal chromatin signature in human embryonic stem cells identifies regulators of cardiac development. Cell. 2012;151(1):221–232.
50.
HeinePDohleEBumsted-O’BrienKEngelkampDSchulteD. Evidence for an evolutionary conserved role of homothorax/Meis1/2 during vertebrate retina development. Development. 2008;135(5):805–811.
51.
ChanRJHromasRYoderMC. The role of Hex in hemangioblast and hematopoietic development. Methods Mol Biol. 2006;330:123–133.
52.
BertolinoEReimundBWildt-PerinicDClercRG. A novel homeobox protein which recognizes a TGT core and functionally interferes with a retinoid-responsive motif. J Biol Chem. 1995;270(52):31178–31188.
53.
BertolinoEWildtSRichardsGClercRG. Expression of a novel murine homeobox gene in the developing cerebellar external granular layer during its proliferation. Dev Dyn. 1996;205(4):410–420.
54.
BaekKBaekJH. The transcription factors myeloid elf-1-like factor (MEF) and distal-less homeobox 5 (Dlx5) inversely regulate the differentiation of osteoblasts and adipocytes in bone marrow. Adipocyte. 2013;2(1):50–54.
