Spheroid formation induces chemokine production in trophoblast-derived Swan71 cells
Tatsuhito Kanda
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorKyosuke Kagami
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorCorresponding Author
Takashi Iizuka
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Correspondence
Hiroshi Fujiwara and Takashi Iizuka, Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa 920-8641, Japan.
Email: [email protected], [email protected] and [email protected]
Search for more papers by this authorHaruki Kasama
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorTakeo Matsumoto
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorYuya Sakai
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorTakuma Suzuki
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorMegumi Yamamoto
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorAyumi Matsuoka
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorRena Yamazaki
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorAkira Hattori
Department of System Chemotherapy and Molecular Sciences, Kyoto University Graduate School of Pharmaceutical Sciences, Kyoto, Japan
Search for more papers by this authorAkihito Horie
Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Kyoto, Japan
Search for more papers by this authorTakiko Daikoku
Division of Animal Disease Model, Research Center for Experimental Modeling of Human Disease, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorMasanori Ono
Department of Obstetrics and Gynecology, Tokyo Medical University, Shinjuku, Tokyo, Japan
Search for more papers by this authorCorresponding Author
Hiroshi Fujiwara
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Correspondence
Hiroshi Fujiwara and Takashi Iizuka, Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa 920-8641, Japan.
Email: [email protected], [email protected] and [email protected]
Search for more papers by this authorTatsuhito Kanda
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorKyosuke Kagami
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorCorresponding Author
Takashi Iizuka
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Correspondence
Hiroshi Fujiwara and Takashi Iizuka, Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa 920-8641, Japan.
Email: [email protected], [email protected] and [email protected]
Search for more papers by this authorHaruki Kasama
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorTakeo Matsumoto
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorYuya Sakai
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorTakuma Suzuki
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorMegumi Yamamoto
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorAyumi Matsuoka
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorRena Yamazaki
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorAkira Hattori
Department of System Chemotherapy and Molecular Sciences, Kyoto University Graduate School of Pharmaceutical Sciences, Kyoto, Japan
Search for more papers by this authorAkihito Horie
Department of Gynecology and Obstetrics, Kyoto University Graduate School of Medicine, Kyoto, Japan
Search for more papers by this authorTakiko Daikoku
Division of Animal Disease Model, Research Center for Experimental Modeling of Human Disease, Kanazawa University, Kanazawa, Ishikawa, Japan
Search for more papers by this authorMasanori Ono
Department of Obstetrics and Gynecology, Tokyo Medical University, Shinjuku, Tokyo, Japan
Search for more papers by this authorCorresponding Author
Hiroshi Fujiwara
Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan
Correspondence
Hiroshi Fujiwara and Takashi Iizuka, Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa 920-8641, Japan.
Email: [email protected], [email protected] and [email protected]
Search for more papers by this authorTatsuhito Kanda and Kyosuke Kagami equally contributed to this study.
Abstract
Problem
In the cell column of anchoring villi, the cytotrophoblast differentiates into extravillous trophoblast (EVT) and invades the endometrium in contact with maternal immune cells. Recently, chemokines were proposed to regulate the decidual immune response. To investigate the roles of chemokines around the anchoring villi, we examined the expression profiles of chemokines in the first-trimester trophoblast-derived Swan71 cells using a three-dimensional culture model.
Method of Study
The gene expressions in the spheroid-formed Swan71 cells were examined by microarray and qPCR analyses. The protein expressions were examined by immunochemical staining. The chemoattractant effects of spheroid-formed Swan71 cells were examined by migration assay using monocyte-derived THP-1 cells.
Results
The expressions of an EVT marker, laeverin, and matrix metalloproteases, MMP2 and MMP9, were increased in the spheroid-cultured Swan71 cells. Microarray and qPCR analysis revealed that mRNA expressions of various chemokines, CCL2, CCL7, CCL20, CXCL1, CXCL2, CXCL5, CXCL6, CXCL8, and CXCL10, in the spheroid-cultured Swan71 cells were up-regulated as compared with those in the monolayer-cultured Swan71 cells. These expressions were significantly suppressed by hypoxia. Migration assay showed that culture media derived from the spheroid-formed Swan71 cells promoted THP-1 cell migration.
Conclusion
This study indicated that chemokine expressions in Swan71 cells increase under a spheroid-forming culture and the culture media have chemoattractant effects. Since three-dimensional cell assembling in the spheroid resembles the structure of the cell column, this study also suggests that chemokines play important roles in the interaction between EVT and immune cells in their early differentiation stage.
CONFLICT OF INTEREST STATEMENT
The authors declare no competing interests.
Open Research
DATA AVAILABILITY STATEMENT
The data that support the findings of this study are available on request from the corresponding author.
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