A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro
Abstract Background Various methods for ex utero culture systems have been explored. However, limitations remain regarding the in vitro culture platforms used before implanting mouse embryos and the normal development of mouse blastocysts in vitro. Furthermore, vascular niche support during mouse em...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-12-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-023-03583-2 |
| _version_ | 1797388482183692288 |
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| author | Junjun Xu Linye Zhang Zihui Ye Binwen Chang Zheng Tu Xuguang Du Xi Wen Yili Teng |
| author_facet | Junjun Xu Linye Zhang Zihui Ye Binwen Chang Zheng Tu Xuguang Du Xi Wen Yili Teng |
| author_sort | Junjun Xu |
| collection | DOAJ |
| description | Abstract Background Various methods for ex utero culture systems have been explored. However, limitations remain regarding the in vitro culture platforms used before implanting mouse embryos and the normal development of mouse blastocysts in vitro. Furthermore, vascular niche support during mouse embryo development from embryonic day (E) 3.5 to E7.5 is unknown in vitro. Methods This study established a three-dimensional (3D) “sandwich” vascular niche culture system with in vitro culture medium (IVCM) using human placenta perivascular stem cells (hPPSCs) and human umbilical vein endothelial cells (hUVECs) as supportive cells (which were seeded into the bottom layer of Matrigel) to test mouse embryos from E3.5 to E7.5 in vitro. The development rates and greatest diameters of mouse embryos from E3.5 to E7.5 were quantitatively determined using SPSS software statistics. Pluripotent markers and embryo transplantation were used to monitor mouse embryo quality and function in vivo. Results Embryos in the IVCM + Cells (hPPSCs + hUVECs) group showed higher development rates and greater diameters at each stage than those in the IVCM group. Embryos in the IVCM + Cells group cultured to E5.5 morphologically resembled natural egg cylinders and expressed specific embryonic cell markers, including Oct4 and Nanog. These features were similar to those of embryos developed in vivo. After transplantation, the embryos were re-implanted in the internal uterus and continued to develop to a particular stage. Conclusions The 3D in vitro culture system enabled embryo development from E3.5 to E7.5, and the vascularization microenvironment constructed by Matrigel, hPPSCs, and hUVECs significantly promoted the development of implanted embryos. This system allowed us to further study the physical and molecular mechanisms of embryo implantation in vitro. |
| first_indexed | 2024-03-08T22:41:28Z |
| format | Article |
| id | doaj.art-83716d36c9d74048b5e86699ca449f96 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-03-08T22:41:28Z |
| publishDate | 2023-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-83716d36c9d74048b5e86699ca449f962023-12-17T12:08:52ZengBMCStem Cell Research & Therapy1757-65122023-12-0114111410.1186/s13287-023-03583-2A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitroJunjun Xu0Linye Zhang1Zihui Ye2Binwen Chang3Zheng Tu4Xuguang Du5Xi Wen6Yili Teng7School of Basic Medical Sciences, Wenzhou Medical UniversityThe First School of Medicine, School of Information and Engineering, The First Affiliated Hospital, Wenzhou Medical UniversityThe First School of Medicine, School of Information and Engineering, The First Affiliated Hospital, Wenzhou Medical UniversitySchool of Public Health and Management, Wenzhou Medical UniversityRenji College, Wenzhou Medical UniversityState Key Laboratory of Animal Biotech Breeding, College of Biological Sciences, China Agricultural UniversityDepartment of Gynecology and Obstetrics, Xuanwu Hospital, Capital Medical UniversityReproductive Medicine Center, The First Affiliated Hospital, Wenzhou Medical UniversityAbstract Background Various methods for ex utero culture systems have been explored. However, limitations remain regarding the in vitro culture platforms used before implanting mouse embryos and the normal development of mouse blastocysts in vitro. Furthermore, vascular niche support during mouse embryo development from embryonic day (E) 3.5 to E7.5 is unknown in vitro. Methods This study established a three-dimensional (3D) “sandwich” vascular niche culture system with in vitro culture medium (IVCM) using human placenta perivascular stem cells (hPPSCs) and human umbilical vein endothelial cells (hUVECs) as supportive cells (which were seeded into the bottom layer of Matrigel) to test mouse embryos from E3.5 to E7.5 in vitro. The development rates and greatest diameters of mouse embryos from E3.5 to E7.5 were quantitatively determined using SPSS software statistics. Pluripotent markers and embryo transplantation were used to monitor mouse embryo quality and function in vivo. Results Embryos in the IVCM + Cells (hPPSCs + hUVECs) group showed higher development rates and greater diameters at each stage than those in the IVCM group. Embryos in the IVCM + Cells group cultured to E5.5 morphologically resembled natural egg cylinders and expressed specific embryonic cell markers, including Oct4 and Nanog. These features were similar to those of embryos developed in vivo. After transplantation, the embryos were re-implanted in the internal uterus and continued to develop to a particular stage. Conclusions The 3D in vitro culture system enabled embryo development from E3.5 to E7.5, and the vascularization microenvironment constructed by Matrigel, hPPSCs, and hUVECs significantly promoted the development of implanted embryos. This system allowed us to further study the physical and molecular mechanisms of embryo implantation in vitro.https://doi.org/10.1186/s13287-023-03583-23D “sandwich” co-culture systemVascular nicheE3.5–E7.5 development |
| spellingShingle | Junjun Xu Linye Zhang Zihui Ye Binwen Chang Zheng Tu Xuguang Du Xi Wen Yili Teng A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro Stem Cell Research & Therapy 3D “sandwich” co-culture system Vascular niche E3.5–E7.5 development |
| title | A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro |
| title_full | A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro |
| title_fullStr | A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro |
| title_full_unstemmed | A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro |
| title_short | A 3D “sandwich” co-culture system with vascular niche supports mouse embryo development from E3.5 to E7.5 in vitro |
| title_sort | 3d sandwich co culture system with vascular niche supports mouse embryo development from e3 5 to e7 5 in vitro |
| topic | 3D “sandwich” co-culture system Vascular niche E3.5–E7.5 development |
| url | https://doi.org/10.1186/s13287-023-03583-2 |
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