Human chorionic gonadotropin from day 2 spent embryo culture media and its relationship to embryo development

A highly sensitive electrochemiluminescence immunoassay for detecting human embryonic human chorionic gonadotropin in spent embryo culture media during IVF-ET cycle

“…2 The concentration of beta-hCG in spent embryo culture media at different stage 1:day1(2PN), 2:day3(5-8 cells), 3:day5(compact/morula), 4: day5(unexpanded blastocyst),5: day5 (expanded blastocyst), 6:day5(arrest) outgrowth embryo. In 2011, beta-hCG was detected in day2 spent embryo culture media by ELISA [4], whose lowest detectable concentration was 1.4 pg/50 μl (0.255mIU/ml), which is within the range from day1 (0.22mIU/ml) to day 3 (0.55mIU/ml) groups in our study. However, these detection methods could only be employed in research due to the limitations such as more sample volume requirement, suboptimal sensitivity and dynamic range.…”

“…Compared with other assays reported before [2][3][4], ECLIA could be automated and did not cause radioactive contamination, besides, its analytical precision (mean intra-assay CV: 4.87 %, mean inter-assay CV: 6.25 %) had been improved considerably, which indicated the high dependability in our study. In addition, there was no significant difference between samples in room temperature and stored at −80°C, further supporting the stability of samples and ECLIA.…”

“…Since embryonic hCG had different isomers in different development stages and current detecting methods could not reach the consistent experimental outcome, it lead to a lack of analysis about the relationship between embryo grading and beta hCG level. According to Sivakumar et al [4], betahCG level from day 2 culture media was independent of the embryo grading in day3 and blastocyst formation. Woodward revealed that only 52 % of the blastocysts could secret hCG, and only 24 % of the original bipronucleate pre-embryos in vitro could be considered anatomically and biochemically competent [10].…”

“…Moreover, beta-hCG was the first reported bio-marker embryonic secretion [2], which is primarily produced by the embryo and later by the syncytiotrophoblast. The hormone was detected at various levels in embryonic culture media from day2 to blastocyst stage by different assays [2][3][4], however, they were only used in research now. An electrochemiluminescence immunoassay (ECLIA) procedure was a method based on solid-phase sandwich immunoassay with advantages of the superior sensitivity of beta-hCG at 0.100mIU/ml in plasma or urine sample and every cycle only took 20 min, which was very popularly employed for diagnosis of conception and trophoblastic disease.…”

Characterization of secreted proteins of 2-cell mouse embryos cultured in vitro to the blastocyst stage with and without protein supplementation

“…Some of these molecules were identified by immunological methods, including among others, paf [20,25], IGF-II [26], HLA-G [27], hCG [28], Apolipoprotein A1 [29] and superoxide dismutase-1 [30]. However, there has been considerable debate as to the reliability of some of these measurements.…”