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| Content Provider | Springer Nature Link |
|---|---|
| Author | Li, Hsin Yang Chang, Sheng Ping Yuan, Chiou Chung Chao, Hsiang Tai Ng, Heung Tat Sung, Yen Jen |
| Copyright Year | 2002 |
| Abstract | During implantation, complex embryo-endometrium interactions result in blastocyst adhesion. To study the mechanisms of implantation, an effective assay for monitoring adhesiveness between embryos and endometrial epithelium is essential. In this study, we describe a simple and reliable method to quantify embryo-endometrium adhesion in vitro. Murine blastocysts or BeWo trophoblast spheroids were cocultured with monolayers of RL95-2 endometrial epithelial cells (EEC) grown in 96-well plates. At the end of coculture, the wells were filled with medium, and the plate was sealed with an adhesive film, inverted, and centrifuged at 25×g for 5 min. After centrifugation, the plate was kept inverted and directly examined microscopically to determine whether the blastocysts or spheroids wereaattached to EEC monolayers. Our assay demonstrated that blastocysts recovered at 1200–1400 h on d 4 were more adherent to EEC than those recovered earlier, consistent with the timing of intrauterine embryo activation. Serum also enhanced blastocyst-EEC adhesion. Spheroid-EEC adhesion was inhibited by blocking Ca$^{2+}$ influx with extracellular Ca$^{2+}$ chelators (ethylenediaminetetraacetic acid or ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid) or a Ca$^{2+}$ channel blocker (verapamil) but not by interfering with Ca$^{2+}$ release from intracellular stores using chelating (1,2-bis(2-aminophenoxy)-ethane-N,N,N′,N′-tetraacetic acid tetrakis(acetoxymethyl ester) or depleting (thapsigargin) agents. Using 96-well plates for coculture, centrifugation, and examination to minimize transfer procedures, our assay system is readily applicable to investigate implantation mechanisms. |
| Starting Page | 505 |
| Ending Page | 511 |
| Page Count | 7 |
| File Format | |
| ISSN | 10712690 |
| Journal | In Vitro Cellular & Developmental Biology - Animal |
| Volume Number | 38 |
| Issue Number | 9 |
| e-ISSN | 1543706X |
| Language | English |
| Publisher | Springer-Verlag |
| Publisher Date | 2002-01-01 |
| Publisher Place | New York |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | adhesion assay blastocyst centrifugation endometrium implantation Cell Biology Developmental Biology Animal Genetics and Genomics Animal Biochemistry Animal Physiology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Developmental Biology Medicine |
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