Detection of IL-6, IL-10, and TNF-α level in human single-blastocyst conditioned medium using ultrasensitive Single Molecule Array platform and its relationship with embryo quality and implantation: a pilot study

J Assist Reprod Genet. 2020 Jul;37(7):1695-1702. doi: 10.1007/s10815-020-01805-7. Epub 2020 May 15.

Abstract

Purpose: The purpose was to investigate the association between embryonic development or implantation and the content of interleukin-6 and 10 (IL-6, IL-10) and tumor necrosis factor-α (TNF-α) in single-blastocyst conditioned medium (SBCM).

Methods: Thirty-eight SBCM samples (SBCMs) were collected from blastocysts with different morphological scores. IL-6, IL-10, and TNF-α concentration in 38 SBCMs was detected by Single Molecule Array and compared according to the blastocyst quality: top-quality (TQ) and non-top quality (NTQ), or blastulation time: day 5 (D5) and day 6 (D6). In another experiment, 61 SBCMs were collected from TQ blastocyst transplanted on D5, and IL-6 concentration in SBCM was compared based on whether embryos are implanted or not (implanted and non-implanted).

Results: In the first experiment, IL-6, IL-10, and TNF-α concentration was not significantly different between the TQ-SBCM and NTQ-SBCM. The D6-SBCM had a higher IL-6 concentration compared with the D5-SBCM, while IL-10 and TNF-α concentration was not significantly different between the D5-SBCM and D6-SBCM. The IL-6 concentration in D5-NTQ or D6-TQ SBCM was higher than that in D5-TQ or D6-NTQ SBCM (P < 0.05), respectively. Furthermore, the spearman analysis demonstrated that IL-6 concentration in SBCM was negatively correlated with the blastocyst quality on D5 and positively correlated with the blastocyst quality on D6. In the second experiment, no significant difference in IL-6 concentration was found between SBCM from implanted and non-implanted blastocyst.

Conclusion: IL-6 concentration in SBCM was associated with embryo quality depending on the blastulation time, although it might not be associated with the blastocyst implantation.

Keywords: Blastocyst quality; Interleukin-10; Interleukin-6; Single Molecule Array platform; Single-blastocyst conditioned medium; Tumor necrosis factor-α.

MeSH terms

  • Adult
  • Biomarkers / analysis
  • Blastocyst / cytology*
  • Blastocyst / metabolism
  • Culture Media, Conditioned / chemistry*
  • Embryo Culture Techniques
  • Embryo Implantation
  • Embryo Transfer
  • Female
  • Humans
  • Interleukin-10 / analysis*
  • Interleukin-6 / analysis*
  • Pilot Projects
  • Pregnancy
  • Pregnancy Rate
  • Single Molecule Imaging / methods
  • Tumor Necrosis Factor-alpha / analysis*

Substances

  • Biomarkers
  • Culture Media, Conditioned
  • IL10 protein, human
  • IL6 protein, human
  • Interleukin-6
  • Tumor Necrosis Factor-alpha
  • Interleukin-10