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Method Article
Lucy Gardner
Centre for Trophoblast Research, University of Cambridge
Corresponding Author
Margherita Turco
Centre for Trophoblast Research, University of Cambridge
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
This protocol describes the generation of long-term, genetically-stable organoid cultures of extraembryonic fetal trophoblasts from human first trimester placentas. Trophoblast organoids are consistently established within one to two weeks. They can be expanded long term, cryopreserved and resurrected. Villous cytotrophoblasts (VCT) expressing EpCam, Ki67 and TP63 form an outer cell layer with deposition of basement membrane in contact with Matrigel. Differentiated, multinucleated syncytial cytotrophoblasts (SCT) form in the centre of the organoid, analogous to the intervillous space. They secrete placental specific peptides and hormones such as hCG, GDF15, PSGs and PAPPA. Characteristic features of SCT are confirmed by electron microscopy (EM). In addition, the organoids can be differentiated to invasive HLA-G+ extravillous trophoblasts (EVT) which digest and migrate out through the Matrigel. In summary, we describe the generation of trophoblast organoids from human extraembryonic tissues that grow as complex 3D structures with fusion of VCT to hCG-secreting SCT, anatomically and functionally closely resembling the villous placenta in vivo.
Keywords
human, placental, trophoblast, organoids, extraembryonic, 3-D culture, Matrigel
The authors declare no competing financial interests.
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Posted 30 Nov, 2018
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Subject Areas
Biological techniques
Associated Publications
Trophoblast organoids as a model for maternal–fetal interactions during human placentation
by Margherita Y. Turco, Lucy Gardner, Richard G. Kay, +15
Nature (27 November, 2018)
Method Article
Lucy Gardner
Centre for Trophoblast Research, University of Cambridge
Corresponding Author
Margherita Turco
Centre for Trophoblast Research, University of Cambridge
Corresponding Author
Version History
CURRENT STATUS: Posted
View the published article here.
Version 1
Posted 30 Nov, 2018
Community comments: 1
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biological techniques
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article here.
This protocol describes the generation of long-term, genetically-stable organoid cultures of extraembryonic fetal trophoblasts from human first trimester placentas. Trophoblast organoids are consistently established within one to two weeks. They can be expanded long term, cryopreserved and resurrected. Villous cytotrophoblasts (VCT) expressing EpCam, Ki67 and TP63 form an outer cell layer with deposition of basement membrane in contact with Matrigel. Differentiated, multinucleated syncytial cytotrophoblasts (SCT) form in the centre of the organoid, analogous to the intervillous space. They secrete placental specific peptides and hormones such as hCG, GDF15, PSGs and PAPPA. Characteristic features of SCT are confirmed by electron microscopy (EM). In addition, the organoids can be differentiated to invasive HLA-G+ extravillous trophoblasts (EVT) which digest and migrate out through the Matrigel. In summary, we describe the generation of trophoblast organoids from human extraembryonic tissues that grow as complex 3D structures with fusion of VCT to hCG-secreting SCT, anatomically and functionally closely resembling the villous placenta in vivo.
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Associated Publications
Trophoblast organoids as a model for maternal–fetal interactions during human placentation
by Margherita Y. Turco, Lucy Gardner, Richard G. Kay, +15
Nature (27 November, 2018)