Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions
IntroductionTrophoblasts are essential in fetal-maternal interaction during pregnancy. The goal was to study HLA profiles of primary trophoblasts derived from placentas, and to investigate their usefulness in studying interaction with immune cells.MethodsAfter enzymatic digestion of first-trimester...
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| Format: | Article |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.814019/full |
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| author | Michael Eikmans Carin van der Keur Jacqueline D. H. Anholts Jos J. M. Drabbels Els van Beelen Susana M. Chuva de Sousa Lopes Marie-Louise van der Hoorn |
| author_facet | Michael Eikmans Carin van der Keur Jacqueline D. H. Anholts Jos J. M. Drabbels Els van Beelen Susana M. Chuva de Sousa Lopes Marie-Louise van der Hoorn |
| author_sort | Michael Eikmans |
| collection | DOAJ |
| description | IntroductionTrophoblasts are essential in fetal-maternal interaction during pregnancy. The goal was to study HLA profiles of primary trophoblasts derived from placentas, and to investigate their usefulness in studying interaction with immune cells.MethodsAfter enzymatic digestion of first-trimester placental tissue from seven donors (6-9 weeks gestation) and trophoblast enrichment we cultured cytotrophoblasts (CTB) in stem cell medium. CTB were differentiated into EVT in a Matrigel-containing medium. A subset of CTB/EVT was profiled for microRNA levels. Expression of classical HLA molecules and of HLA-G was studied by flow cytometry, qPCR, and ELISA. Secondary trophoblast cell lines JAR and JEG-3 were studied as controls. Lymphocytes were investigated during co-culturing with EVT.ResultsThe trophoblasts could be easily maintained for several passages, upregulated classical trophoblast markers (GATA3, TFAP2C, chromosome-19 microRNAs), and upon differentiation to EVT they were selective in expressing HLA-C. EVT showed increasing expression of total HLA-G, an increasing proportion of HLA-G1 over G2- and G3 isoforms, and elevated excretion of soluble HLA-G. These features were distinct from those of the secondary trophoblast cell lines. TNF-α and IL-8 represented the most abundantly secreted cytokines by CTB, but their levels were minimal in EVT cultures. As proof of principle, we showed that EVT affect lymphocytes in three-day co-cultures (n=4) by decreasing activation marker HLA-DR.ConclusionWe verified the possibility culturing trophoblasts from first-term placentas, and their capability of differentiating to HLA-G expressing EVT. This culture model better represents the in-vivo situation than previously studied secondary trophoblast cell lines and enables mechanistic studies of fetal-maternal interactions. |
| first_indexed | 2024-12-12T03:31:35Z |
| format | Article |
| id | doaj.art-9e51ceb2587f4c7897a158cb971dcea0 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-12T03:31:35Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-9e51ceb2587f4c7897a158cb971dcea02022-12-22T00:39:54ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-05-011310.3389/fimmu.2022.814019814019Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell InteractionsMichael Eikmans0Carin van der Keur1Jacqueline D. H. Anholts2Jos J. M. Drabbels3Els van Beelen4Susana M. Chuva de Sousa Lopes5Marie-Louise van der Hoorn6Department of Immunology, Leiden University Medical Center, Leiden, NetherlandsDepartment of Immunology, Leiden University Medical Center, Leiden, NetherlandsDepartment of Immunology, Leiden University Medical Center, Leiden, NetherlandsDepartment of Immunology, Leiden University Medical Center, Leiden, NetherlandsDepartment of Immunology, Leiden University Medical Center, Leiden, NetherlandsDepartment of Anatomy and Embryology, Leiden University Medical Center, Leiden, NetherlandsDepartment of Gynecology and Obstetrics, Leiden University Medical Center, Leiden, NetherlandsIntroductionTrophoblasts are essential in fetal-maternal interaction during pregnancy. The goal was to study HLA profiles of primary trophoblasts derived from placentas, and to investigate their usefulness in studying interaction with immune cells.MethodsAfter enzymatic digestion of first-trimester placental tissue from seven donors (6-9 weeks gestation) and trophoblast enrichment we cultured cytotrophoblasts (CTB) in stem cell medium. CTB were differentiated into EVT in a Matrigel-containing medium. A subset of CTB/EVT was profiled for microRNA levels. Expression of classical HLA molecules and of HLA-G was studied by flow cytometry, qPCR, and ELISA. Secondary trophoblast cell lines JAR and JEG-3 were studied as controls. Lymphocytes were investigated during co-culturing with EVT.ResultsThe trophoblasts could be easily maintained for several passages, upregulated classical trophoblast markers (GATA3, TFAP2C, chromosome-19 microRNAs), and upon differentiation to EVT they were selective in expressing HLA-C. EVT showed increasing expression of total HLA-G, an increasing proportion of HLA-G1 over G2- and G3 isoforms, and elevated excretion of soluble HLA-G. These features were distinct from those of the secondary trophoblast cell lines. TNF-α and IL-8 represented the most abundantly secreted cytokines by CTB, but their levels were minimal in EVT cultures. As proof of principle, we showed that EVT affect lymphocytes in three-day co-cultures (n=4) by decreasing activation marker HLA-DR.ConclusionWe verified the possibility culturing trophoblasts from first-term placentas, and their capability of differentiating to HLA-G expressing EVT. This culture model better represents the in-vivo situation than previously studied secondary trophoblast cell lines and enables mechanistic studies of fetal-maternal interactions.https://www.frontiersin.org/articles/10.3389/fimmu.2022.814019/fulltrophoblastHLA-Gplacentapregnancyculturingstem cell |
| spellingShingle | Michael Eikmans Carin van der Keur Jacqueline D. H. Anholts Jos J. M. Drabbels Els van Beelen Susana M. Chuva de Sousa Lopes Marie-Louise van der Hoorn Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions Frontiers in Immunology trophoblast HLA-G placenta pregnancy culturing stem cell |
| title | Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions |
| title_full | Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions |
| title_fullStr | Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions |
| title_full_unstemmed | Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions |
| title_short | Primary Trophoblast Cultures: Characterization of HLA Profiles and Immune Cell Interactions |
| title_sort | primary trophoblast cultures characterization of hla profiles and immune cell interactions |
| topic | trophoblast HLA-G placenta pregnancy culturing stem cell |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.814019/full |
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