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J Cell Biol. 1989 August 1; 109(2): 891–902.
PMCID: PMC2115717

Adhesive and degradative properties of human placental cytotrophoblast cells in vitro


Human fetal development depends on the embryo rapidly gaining access to the maternal circulation. The trophoblast cells that form the fetal portion of the human placenta have solved this problem by transiently exhibiting certain tumor-like properties. Thus, during early pregnancy fetal cytotrophoblast cells invade the uterus and its arterial network. This process peaks during the twelfth week of pregnancy and declines rapidly thereafter, suggesting that the highly specialized, invasive behavior of the cytotrophoblast cells is closely regulated. Since little is known about the actual mechanisms involved, we developed an isolation procedure for cytotrophoblasts from placentas of different gestational ages to study their adhesive and invasive properties in vitro. Cytotrophoblasts isolated from first, second, and third trimester human placentas were plated on the basement membrane-like extracellular matrix produced by the PF HR9 teratocarcinoma cell line. Cells from all trimesters expressed the calcium-dependent cell adhesion molecule cell-CAM 120/80 (E-cadherin) which, in the placenta, is specific for cytotrophoblasts. However, only the first trimester cytotrophoblast cells degraded the matrices on which they were cultured, leaving large gaps in the basement membrane substrates and releasing low molecular mass 3H-labeled matrix components into the medium. No similar degradative activity was observed when second or third trimester cytotrophoblast cells, first trimester human placental fibroblasts, or the human choriocarcinoma cell lines BeWo and JAR were cultured on radiolabeled matrices. To begin to understand the biochemical basis of this degradative behavior, the substrate gel technique was used to analyze the cell-associated and secreted proteinase activities expressed by early, mid, and late gestation cytotrophoblasts. Several gelatin-degrading proteinases were uniquely expressed by early gestation, invasive cytotrophoblasts, and all these activities could be abolished by inhibitors of metalloproteinases. By early second trimester, the time when cytotrophoblast invasion rapidly diminishes in vivo, the proteinase pattern of the cytotrophoblasts was identical to that of term, noninvasive cells. These results are the first evidence suggesting that specialized, temporally regulated metalloproteinases are involved in trophoblast invasion of the uterus. Since the cytotrophoblasts from first trimester and later gestation placentas maintain for several days the temporally regulated degradative behavior displayed in vivo, the short-term cytotrophoblast outgrowth culture system described here should be useful in studying some of the early events in human placen

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Selected References

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  • Blackwood CE, Hosannah Y, Mandl I. Proteolytic enzyme systems in developing rat tissues. J Reprod Fertil. 1968 Oct;17(1):19–33. [PubMed]
  • Brosens I, Dixon HG. The anatomy of the maternal side of the placenta. J Obstet Gynaecol Br Commonw. 1966 Jun;73(3):357–363. [PubMed]
  • Chin JR, Murphy G, Werb Z. Stromelysin, a connective tissue-degrading metalloendopeptidase secreted by stimulated rabbit synovial fibroblasts in parallel with collagenase. Biosynthesis, isolation, characterization, and substrates. J Biol Chem. 1985 Oct 5;260(22):12367–12376. [PubMed]
  • Cotte C, Easty GC, Neville AM, Monaghan P. Preparation of highly purified cytotrophoblast from human placenta with subsequent modulation to form syncytiotrophoblast in monolayer cultures. In Vitro. 1980 Aug;16(8):639–646. [PubMed]
  • Damsky CH, Richa J, Solter D, Knudsen K, Buck CA. Identification and purification of a cell surface glycoprotein mediating intercellular adhesion in embryonic and adult tissue. Cell. 1983 Sep;34(2):455–466. [PubMed]
  • Damsky CH, Knudsen KA, Bradley D, Buck CA, Horwitz AF. Distribution of the cell substratum attachment (CSAT) antigen on myogenic and fibroblastic cells in culture. J Cell Biol. 1985 May;100(5):1528–1539. [PMC free article] [PubMed]
  • Denker HW. Implantation. The role of proteinases, and blockage of implantation by proteinase inhibitors. Adv Anat Embryol Cell Biol. 1977;53(5):3–123. [PubMed]
  • Denker HW, Fritz H. Enzymic characterization of rabbit blastocyst proteinase with synthetic substrates of trypsin-like enzymes. Hoppe Seylers Z Physiol Chem. 1979 Feb;360(2):107–113. [PubMed]
  • DeVilla GO, Jr, Roberts K, Wiest WG, Mikhail G, Flickinger G. A specific radioimmunoassay of plasma progesterone. J Clin Endocrinol Metab. 1972 Sep;35(3):458–460. [PubMed]
  • Enders AC. Cytology of human early implantation. Res Reprod. 1976 Sep;8(5):1–2. [PubMed]
  • Fisher SJ, Leitch MS, Kantor MS, Basbaum CB, Kramer RH. Degradation of extracellular matrix by the trophoblastic cells of first-trimester human placentas. J Cell Biochem. 1985;27(1):31–41. [PubMed]
  • Gallin WJ, Edelman GM, Cunningham BA. Characterization of L-CAM, a major cell adhesion molecule from embryonic liver cells. Proc Natl Acad Sci U S A. 1983 Feb;80(4):1038–1042. [PubMed]
  • Glass RH, Spindle AI, Pedersen RA. Mouse embryo attachment to substratum and interaction of trophoblast with cultured cells. J Exp Zool. 1979 Jun;208(3):327–336. [PubMed]
  • Glass RH, Aggeler J, Spindle A, Pedersen RA, Werb Z. Degradation of extracellular matrix by mouse trophoblast outgrowths: a model for implantation. J Cell Biol. 1983 Apr;96(4):1108–1116. [PMC free article] [PubMed]
  • Gwatkin RB. Defined media and development of mammalian eggs in vitro. Ann N Y Acad Sci. 1966 Oct 7;139(1):79–90. [PubMed]
  • HERTIG AT, ROCK J, ADAMS EC. A description of 34 human ova within the first 17 days of development. Am J Anat. 1956 May;98(3):435–493. [PubMed]
  • Heussen C, Dowdle EB. Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Anal Biochem. 1980 Feb;102(1):196–202. [PubMed]
  • Kliman HJ, Nestler JE, Sermasi E, Sanger JM, Strauss JF., 3rd Purification, characterization, and in vitro differentiation of cytotrophoblasts from human term placentae. Endocrinology. 1986 Apr;118(4):1567–1582. [PubMed]
  • Kubo H, Spindle A, Pedersen RA. Inhibition of mouse blastocyst attachment and outgrowth by protease inhibitors. J Exp Zool. 1981 Jun;216(3):445–451. [PubMed]
  • Lindenberg S, Hyttel P, Lenz S, Holmes PV. Ultrastructure of the early human implantation in vitro. Hum Reprod. 1986 Dec;1(8):533–538. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • McKerrow JH, Pino-Heiss S, Lindquist R, Werb Z. Purification and characterization of an elastinolytic proteinase secreted by cercariae of Schistosoma mansoni. J Biol Chem. 1985 Mar 25;260(6):3703–3707. [PubMed]
  • Nelson DM, Meister RK, Ortman-Nabi J, Sparks S, Stevens VC. Differentiation and secretory activities of cultured human placental cytotrophoblast. Placenta. 1986 Jan-Feb;7(1):1–16. [PubMed]
  • Nilsson O. The morphology of blastocyst implantation. J Reprod Fertil. 1974 Jul;39(1):187–194. [PubMed]
  • Pattillo RA, Gey GO. The establishment of a cell line of human hormone-synthesizing trophoblastic cells in vitro. Cancer Res. 1968 Jul;28(7):1231–1236. [PubMed]
  • Peters D, Branscomb E, Dean P, Merrill T, Pinkel D, Van Dilla M, Gray JW. The LLNL high-speed sorter: design features, operational characteristics, and biological utility. Cytometry. 1985 Jul;6(4):290–301. [PubMed]
  • Peyriéras N, Hyafil F, Louvard D, Ploegh HL, Jacob F. Uvomorulin: a nonintegral membrane protein of early mouse embryo. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6274–6277. [PubMed]
  • Queenan JT, Jr, Kao LC, Arboleda CE, Ulloa-Aguirre A, Golos TG, Cines DB, Strauss JF., 3rd Regulation of urokinase-type plasminogen activator production by cultured human cytotrophoblasts. J Biol Chem. 1987 Aug 15;262(23):10903–10906. [PubMed]
  • Ramsey EM, Houston ML, Harris JW. Interactions of the trophoblast and maternal tissues in three closely related primate species. Am J Obstet Gynecol. 1976 Mar 15;124(6):647–652. [PubMed]
  • Reich R, Thompson EW, Iwamoto Y, Martin GR, Deason JR, Fuller GC, Miskin R. Effects of inhibitors of plasminogen activator, serine proteinases, and collagenase IV on the invasion of basement membranes by metastatic cells. Cancer Res. 1988 Jun 15;48(12):3307–3312. [PubMed]
  • Robertson WB, Brosens I, Dixon G. Uteroplacental vascular pathology. Eur J Obstet Gynecol Reprod Biol. 1975;5(1-2):47–65. [PubMed]
  • Salomon DS, Sherman MI. Implantation and invasiveness of mouse blastocysts on uterine monolayers. Exp Cell Res. 1975 Feb;90(2):261–268. [PubMed]
  • Sherman MI. Long term culture of cells derived from mouse blastocysts. Differentiation. 1975 Aug 11;3(1-3):51–67. [PubMed]
  • Sherman MI, Barlow PW. Deoxyribonucleic acid content in delayed mouse blastocysts. J Reprod Fertil. 1972 Apr;29(1):123–126. [PubMed]
  • Sherman MI, Salomon DS. The relationships between the early mouse embryo and its environment. Symp Soc Dev Biol. 1975;(33):277–309. [PubMed]
  • Spindle AI, Pedersen RA. Hatching, attachment, and outgrowth of mouse blastocysts in vitro: fixed nitrogen requirements. J Exp Zool. 1973 Dec;186(3):305–318. [PubMed]
  • Strickland S, Beers WH. Studies on the role of plasminogen activator in ovulation. In vitro response of granulosa cells to gonadotropins, cyclic nucleotides, and prostaglandins. J Biol Chem. 1976 Sep 25;251(18):5694–5702. [PubMed]
  • Strickland S, Reich E, Sherman MI. Plasminogen activator in early embryogenesis: enzyme production by trophoblast and parietal endoderm. Cell. 1976 Oct;9(2):231–240. [PubMed]
  • Takeichi M. The cadherins: cell-cell adhesion molecules controlling animal morphogenesis. Development. 1988 Apr;102(4):639–655. [PubMed]
  • Tuttle SE, O'Toole RV, O'Shaughnessy RW, Zuspan FP. Immunohistochemical evaluation of human placental implantation: an initial study. Am J Obstet Gynecol. 1985 Oct 1;153(3):239–244. [PubMed]
  • Unkeless JC, Gordon S, Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. [PMC free article] [PubMed]
  • Wewer UM, Faber M, Liotta LA, Albrechtsen R. Immunochemical and ultrastructural assessment of the nature of the pericellular basement membrane of human decidual cells. Lab Invest. 1985 Dec;53(6):624–633. [PubMed]
  • Wewer UM, Damjanov A, Weiss J, Liotta LA, Damjanov I. Mouse endometrial stromal cells produce basement-membrane components. Differentiation. 1986;32(1):49–58. [PubMed]
  • Wheelock MJ, Buck CA, Bechtol KB, Damsky CH. Soluble 80-kd fragment of cell-CAM 120/80 disrupts cell-cell adhesion. J Cell Biochem. 1987 Jul;34(3):187–202. [PubMed]
  • Yagel S, Parhar RS, Jeffrey JJ, Lala PK. Normal nonmetastatic human trophoblast cells share in vitro invasive properties of malignant cells. J Cell Physiol. 1988 Sep;136(3):455–462. [PubMed]
  • Yoshida-Noro C, Suzuki N, Takeichi M. Molecular nature of the calcium-dependent cell-cell adhesion system in mouse teratocarcinoma and embryonic cells studied with a monoclonal antibody. Dev Biol. 1984 Jan;101(1):19–27. [PubMed]

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