Trophoblast differentiation, invasion and hormone secretion in a three-dimensional in vitro implantation model with rhesus monkey embryos

Tiencheng A Chang, Gennadiy I. Bondarenko, Behzad Gerami-Naini, Jessica G. Drenzek, Maureen Durning, Mark A. Garthwaite, Jenna Kropp Schmidt, Thaddeus G. Golos

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: The initiation of primate embryo invasion into the endometrium and the formation of the placenta from trophoblasts, fetal mesenchyme, and vascular components are essential for the establishment of a successful pregnancy. The mechanisms which direct morphogenesis of the chorionic villi, and the interactions between trophectoderm-derived trophoblasts and the fetal mesenchyme to direct these processes during placentation are not well understood due to a dearth of systems to examine and manipulate real-time primate implantation. Here we describe an in vitro three-dimensional (3-D) model to study implantation which utilized IVF-generated rhesus monkey embryos cultured in a Matrigel explant system. Methods: Blastocyst stage embryos were embedded in a 3-D microenvironment of a Matrigel carrier and co-cultured with a feeder layer of cells generating conditioned medium. Throughout the course of embryo co-culture embryo growth and secretions were monitored. Embedded embryos were then sectioned and stained for markers of trophoblast function and differentiation. Results: Signs of implantation were observed including enlargement of the embryo mass, and invasion and proliferation of trophoblast outgrowths. Expression of chorionic gonadotropin defined by immunohistochemical staining, and secretion of chorionic gonadotropin and progesterone coincident with the appearance of trophoblast outgrowths, supported the conclusion that a trophoblast cell lineage formed from implanted embryos. Positive staining for selected markers including Ki67, MHC class I, NeuN, CD31, vonWillebrand Factor and Vimentin, suggest growth and differentiation of the embryo following embedding. Conclusions: This 3-D in vitro system will facilitate further study of primate embryo biology, with potential to provide a platform for study of genes related to implantation defects and trophoblast differentiation.

Original languageEnglish (US)
Article number24
JournalReproductive Biology and Endocrinology
Volume16
Issue number1
DOIs
StatePublished - Mar 16 2018

Fingerprint

Trophoblasts
Macaca mulatta
Embryonic Structures
Hormones
Primates
Mesoderm
Chorionic Gonadotropin
In Vitro Techniques
Staining and Labeling
Feeder Cells
Chorionic Villi
Placentation
Blastocyst
Vimentin
Cell Lineage
Conditioned Culture Medium
Growth
Endometrium
Coculture Techniques
Morphogenesis

Keywords

  • Embryo
  • Implantation
  • Non-human primate
  • Trophoblast

ASJC Scopus subject areas

  • Reproductive Medicine
  • Endocrinology
  • Obstetrics and Gynecology
  • Developmental Biology

Cite this

Trophoblast differentiation, invasion and hormone secretion in a three-dimensional in vitro implantation model with rhesus monkey embryos. / Chang, Tiencheng A; Bondarenko, Gennadiy I.; Gerami-Naini, Behzad; Drenzek, Jessica G.; Durning, Maureen; Garthwaite, Mark A.; Schmidt, Jenna Kropp; Golos, Thaddeus G.

In: Reproductive Biology and Endocrinology, Vol. 16, No. 1, 24, 16.03.2018.

Research output: Contribution to journalArticle

Chang, Tiencheng A ; Bondarenko, Gennadiy I. ; Gerami-Naini, Behzad ; Drenzek, Jessica G. ; Durning, Maureen ; Garthwaite, Mark A. ; Schmidt, Jenna Kropp ; Golos, Thaddeus G. / Trophoblast differentiation, invasion and hormone secretion in a three-dimensional in vitro implantation model with rhesus monkey embryos. In: Reproductive Biology and Endocrinology. 2018 ; Vol. 16, No. 1.
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abstract = "Background: The initiation of primate embryo invasion into the endometrium and the formation of the placenta from trophoblasts, fetal mesenchyme, and vascular components are essential for the establishment of a successful pregnancy. The mechanisms which direct morphogenesis of the chorionic villi, and the interactions between trophectoderm-derived trophoblasts and the fetal mesenchyme to direct these processes during placentation are not well understood due to a dearth of systems to examine and manipulate real-time primate implantation. Here we describe an in vitro three-dimensional (3-D) model to study implantation which utilized IVF-generated rhesus monkey embryos cultured in a Matrigel explant system. Methods: Blastocyst stage embryos were embedded in a 3-D microenvironment of a Matrigel carrier and co-cultured with a feeder layer of cells generating conditioned medium. Throughout the course of embryo co-culture embryo growth and secretions were monitored. Embedded embryos were then sectioned and stained for markers of trophoblast function and differentiation. Results: Signs of implantation were observed including enlargement of the embryo mass, and invasion and proliferation of trophoblast outgrowths. Expression of chorionic gonadotropin defined by immunohistochemical staining, and secretion of chorionic gonadotropin and progesterone coincident with the appearance of trophoblast outgrowths, supported the conclusion that a trophoblast cell lineage formed from implanted embryos. Positive staining for selected markers including Ki67, MHC class I, NeuN, CD31, vonWillebrand Factor and Vimentin, suggest growth and differentiation of the embryo following embedding. Conclusions: This 3-D in vitro system will facilitate further study of primate embryo biology, with potential to provide a platform for study of genes related to implantation defects and trophoblast differentiation.",
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AU - Chang, Tiencheng A

AU - Bondarenko, Gennadiy I.

AU - Gerami-Naini, Behzad

AU - Drenzek, Jessica G.

AU - Durning, Maureen

AU - Garthwaite, Mark A.

AU - Schmidt, Jenna Kropp

AU - Golos, Thaddeus G.

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AB - Background: The initiation of primate embryo invasion into the endometrium and the formation of the placenta from trophoblasts, fetal mesenchyme, and vascular components are essential for the establishment of a successful pregnancy. The mechanisms which direct morphogenesis of the chorionic villi, and the interactions between trophectoderm-derived trophoblasts and the fetal mesenchyme to direct these processes during placentation are not well understood due to a dearth of systems to examine and manipulate real-time primate implantation. Here we describe an in vitro three-dimensional (3-D) model to study implantation which utilized IVF-generated rhesus monkey embryos cultured in a Matrigel explant system. Methods: Blastocyst stage embryos were embedded in a 3-D microenvironment of a Matrigel carrier and co-cultured with a feeder layer of cells generating conditioned medium. Throughout the course of embryo co-culture embryo growth and secretions were monitored. Embedded embryos were then sectioned and stained for markers of trophoblast function and differentiation. Results: Signs of implantation were observed including enlargement of the embryo mass, and invasion and proliferation of trophoblast outgrowths. Expression of chorionic gonadotropin defined by immunohistochemical staining, and secretion of chorionic gonadotropin and progesterone coincident with the appearance of trophoblast outgrowths, supported the conclusion that a trophoblast cell lineage formed from implanted embryos. Positive staining for selected markers including Ki67, MHC class I, NeuN, CD31, vonWillebrand Factor and Vimentin, suggest growth and differentiation of the embryo following embedding. Conclusions: This 3-D in vitro system will facilitate further study of primate embryo biology, with potential to provide a platform for study of genes related to implantation defects and trophoblast differentiation.

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