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Article: Establishment of a novel human embryonic stem cell-derived trophoblastic spheroid implantation model
Title | Establishment of a novel human embryonic stem cell-derived trophoblastic spheroid implantation model |
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Authors | |
Keywords | embryo development human endometrium implantation in vitro implantation model stem cells trophoblasts |
Issue Date | 2015 |
Publisher | Oxford University Press. The Journal's web site is located at http://humrep.oxfordjournals.org/ |
Citation | Human Reproduction, 2015, v. 30 n. 11, p. 2614-2626 How to Cite? |
Abstract | STUDY QUESTION Can human embryonic stem cell-derived trophoblastic spheroids be used to study the early stages of implantation? SUMMARY ANSWER We generated a novel human embryonic stem cell-derived trophoblastic spheroid model mimicking human blastocysts in the early stages of implantation. WHAT IS KNOWN ALREADY Both human embryos and choriocarcinoma cell line derived spheroids can attach onto endometrial cells and are used as models to study the early stages of implantation. However, human embryos are limited and the use of cancer cell lines for spheroid generation remains sub-optimal for research. STUDY DESIGN, SIZE, DURATION Experimental induced differentiation of human embryonic stem cells into trophoblast and characterization of the trophoblast. PARTICIPANTS/MATERIALS, SETTING, METHODS Trophoblastic spheroids (BAP-EB) were generated by inducing differentiation of a human embryonic stem cell line, VAL3 cells with bone morphogenic factor-4, A83-01 (a TGF-β inhibitor), and PD173074 (a FGF receptor-3 inhibitor) after embryoid body formation. The expressions of trophoblastic markers and hCG levels were studied by real-time PCR and immunohistochemistry. BAP-EB attachment and invasion assays were performed on different cell lines and primary endometrial cells. MAIN RESULTS AND THE ROLE OF CHANCE After 48 h of induced differentiation, the BAP-EB resembled early implanting human embryos in terms of size and morphology. The spheroids derived from embryonic stem cells (VAL3), but not from several other cell lines studied, possessed a blastocoel-like cavity. BAP-EB expressed several markers of trophectoderm of human blastocysts on Day 2 of induced differentiation. In the subsequent days of differentiation, the cells of the spheroids differentiated into trophoblast-like cells expressing trophoblastic markers, though at levels lower than that in the primary trophoblasts or in a choriocarcinoma cell line. On Day 3 of induced differentiation, BAP-EB selectively attached onto endometrial epithelial cells, but not other non-endometrial cell lines or an endometrial cell line that had lost its epithelial character. The attachment rates of BAP-EB was significantly higher on primary endometrial epithelial cells (EEC) taken from 7 days after hCG induction of ovulation (hCG+7 day) when compared with that from hCG+2 day. The spheroids also invaded through Ishikawa cells and the primary endometrial stromal cells in the co-culture. LIMITATIONS, REASONS FOR CAUTION The attachment rates of BAP-EB were compared between EEC obtained from Day 2 and Day 7 of the gonadotrophin stimulated cycle, but not the natural cycles. WIDER IMPLICATIONS OF THE FINDINGS BAP-EB have the potential to be used as a test for predicting endometrial receptivity in IVF cycles and provide a novel approach to study early human implantation, trophoblastic cell differentiation and trophoblastic invasion into human endometrial cells. STUDY FUNDING/COMPETING INTEREST(S) This study was supported in part by a General Research Fund (grant number: 17111414) from the Research Grants Council of Hong Kong. The authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER Nil. © 2015 The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. |
Persistent Identifier | http://hdl.handle.net/10722/227475 |
ISSN | 2023 Impact Factor: 6.0 2023 SCImago Journal Rankings: 1.852 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Lee, CYL | - |
dc.contributor.author | Fong, SW | - |
dc.contributor.author | Chen, ACH | - |
dc.contributor.author | Li, T | - |
dc.contributor.author | Yue, C | - |
dc.contributor.author | Lee, CL | - |
dc.contributor.author | Ng, EHY | - |
dc.contributor.author | Yeung, WSB | - |
dc.contributor.author | Lee, CKF | - |
dc.date.accessioned | 2016-07-18T09:10:56Z | - |
dc.date.available | 2016-07-18T09:10:56Z | - |
dc.date.issued | 2015 | - |
dc.identifier.citation | Human Reproduction, 2015, v. 30 n. 11, p. 2614-2626 | - |
dc.identifier.issn | 0268-1161 | - |
dc.identifier.uri | http://hdl.handle.net/10722/227475 | - |
dc.description.abstract | STUDY QUESTION Can human embryonic stem cell-derived trophoblastic spheroids be used to study the early stages of implantation? SUMMARY ANSWER We generated a novel human embryonic stem cell-derived trophoblastic spheroid model mimicking human blastocysts in the early stages of implantation. WHAT IS KNOWN ALREADY Both human embryos and choriocarcinoma cell line derived spheroids can attach onto endometrial cells and are used as models to study the early stages of implantation. However, human embryos are limited and the use of cancer cell lines for spheroid generation remains sub-optimal for research. STUDY DESIGN, SIZE, DURATION Experimental induced differentiation of human embryonic stem cells into trophoblast and characterization of the trophoblast. PARTICIPANTS/MATERIALS, SETTING, METHODS Trophoblastic spheroids (BAP-EB) were generated by inducing differentiation of a human embryonic stem cell line, VAL3 cells with bone morphogenic factor-4, A83-01 (a TGF-β inhibitor), and PD173074 (a FGF receptor-3 inhibitor) after embryoid body formation. The expressions of trophoblastic markers and hCG levels were studied by real-time PCR and immunohistochemistry. BAP-EB attachment and invasion assays were performed on different cell lines and primary endometrial cells. MAIN RESULTS AND THE ROLE OF CHANCE After 48 h of induced differentiation, the BAP-EB resembled early implanting human embryos in terms of size and morphology. The spheroids derived from embryonic stem cells (VAL3), but not from several other cell lines studied, possessed a blastocoel-like cavity. BAP-EB expressed several markers of trophectoderm of human blastocysts on Day 2 of induced differentiation. In the subsequent days of differentiation, the cells of the spheroids differentiated into trophoblast-like cells expressing trophoblastic markers, though at levels lower than that in the primary trophoblasts or in a choriocarcinoma cell line. On Day 3 of induced differentiation, BAP-EB selectively attached onto endometrial epithelial cells, but not other non-endometrial cell lines or an endometrial cell line that had lost its epithelial character. The attachment rates of BAP-EB was significantly higher on primary endometrial epithelial cells (EEC) taken from 7 days after hCG induction of ovulation (hCG+7 day) when compared with that from hCG+2 day. The spheroids also invaded through Ishikawa cells and the primary endometrial stromal cells in the co-culture. LIMITATIONS, REASONS FOR CAUTION The attachment rates of BAP-EB were compared between EEC obtained from Day 2 and Day 7 of the gonadotrophin stimulated cycle, but not the natural cycles. WIDER IMPLICATIONS OF THE FINDINGS BAP-EB have the potential to be used as a test for predicting endometrial receptivity in IVF cycles and provide a novel approach to study early human implantation, trophoblastic cell differentiation and trophoblastic invasion into human endometrial cells. STUDY FUNDING/COMPETING INTEREST(S) This study was supported in part by a General Research Fund (grant number: 17111414) from the Research Grants Council of Hong Kong. The authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER Nil. © 2015 The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. | - |
dc.language | eng | - |
dc.publisher | Oxford University Press. The Journal's web site is located at http://humrep.oxfordjournals.org/ | - |
dc.relation.ispartof | Human Reproduction | - |
dc.rights | This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Human Reproduction following peer review. The definitive publisher-authenticated version Human Reproduction, 2015, v. 30 n. 11, p. 2614-2626 is available online at: http://humrep.oxfordjournals.org/content/30/11/2614 | - |
dc.subject | embryo development | - |
dc.subject | human endometrium | - |
dc.subject | implantation | - |
dc.subject | in vitro implantation model | - |
dc.subject | stem cells | - |
dc.subject | trophoblasts | - |
dc.title | Establishment of a novel human embryonic stem cell-derived trophoblastic spheroid implantation model | - |
dc.type | Article | - |
dc.identifier.email | Lee, CYL: cherielee@hku.hk | - |
dc.identifier.email | Fong, SW: szewan11@hku.hk | - |
dc.identifier.email | Chen, ACH: andycch0@hku.hk | - |
dc.identifier.email | Lee, CL: kcllee@hku.hk | - |
dc.identifier.email | Ng, EHY: nghye@hku.hk | - |
dc.identifier.email | Yeung, WSB: wsbyeung@hku.hk | - |
dc.identifier.email | Lee, CKF: ckflee@hku.hk | - |
dc.identifier.authority | Lee, CYL=rp00308 | - |
dc.identifier.authority | Ng, EHY=rp00426 | - |
dc.identifier.authority | Yeung, WSB=rp00331 | - |
dc.identifier.authority | Lee, CKF=rp00458 | - |
dc.description.nature | postprint | - |
dc.identifier.doi | 10.1093/humrep/dev223 | - |
dc.identifier.pmid | 26350609 | - |
dc.identifier.scopus | eid_2-s2.0-84946415560 | - |
dc.identifier.hkuros | 258957 | - |
dc.identifier.volume | 30 | - |
dc.identifier.issue | 11 | - |
dc.identifier.spage | 2614 | - |
dc.identifier.epage | 2626 | - |
dc.identifier.isi | WOS:000363053400019 | - |
dc.publisher.place | United Kingdom | - |
dc.identifier.issnl | 0268-1161 | - |