Potential of inner cell mass outgrowth and amino acid turnover as markers of quality in the in vitro fertilization laboratory

Capsule:
Inner cell mass outgrowth is a more sensitive bioassay than blastocyst development and sperm motility for detecting formaldehyde. Evaluation of culture media metabolome has potential as a sensitive QC assay.

Authors:
Ravi P. Gada, M.D., Gaurang S. Daftary, M.D., David L. Walker, M.S., Jean M. Lacey, B.A., Dietrich Matern, M.D., Ph.D., Dean E. Morbeck, Ph.D.

Volume 98, Issue 4, Pages 863-869.e1, October 2012

Abstract:

Objective:
To compare sensitivity of inner cell mass (ICM) outgrowth assay and analysis of culture media amino acid turnover to the sensitivity of the human sperm motility assay (HSMA) and murine embryo assay (MEA) for detection of formaldehyde toxicity.

Design:
Prospective in vitro study.

Settings:
University hospital-based infertility center.

Animal(s):
Murine embryos.

Intervention:
HSMA, MEA, and ICM outgrowth assays were performed with media containing 0–64 uM concentrations of formaldehyde. These assays were compared to dynamics of amino acid turnover in culture media.

Main Outcome Measure(s):
The lowest concentration of formaldehyde in culture media detected by each quality control assay.

Result(s):
Sperm forward progression, but not motility, detected formaldehyde at a concentration of 32 uM. Sperm motility index identified formaldehyde toxicity at 64 uM whereas blastocyst rates in the MEA were affected at 32 uM formaldehyde. Evaluation of ICM using outgrowth and grade detected 16 uM formaldehyde. Leucine turnover in culture media detected 64 uM formaldehyde in the amino acid assay.

Conclusion(s):
Inner cell mass outgrowth is a more sensitive bioassay than MEA and HSMA for the detection of formaldehyde in culture media. Amino acid metabolism may also provide a sensitive quality control measure for detection of formaldehyde.

  • Maria Jose De Los Santos

    I like very much the idea to go further and test the ICM outgrowth as a new QC strategy. I think most of the clinical embryologyst feel that that the current QC assays are not sensitive enough and new aproaches are needed. It would be very nice to have human blactocyts donated to research and see whether a correlation exists with this mouse based model.

    • In our lab, we feel the same way. Most labs can get a mouse embryo to develop to a blastocyst in poor culture conditions. The culture beyond that to ICM outgrowth requires better conditions. Thank you for the comment.

  • Thank you Micah. In terms of cost it is not that much extra. It is more time intensive, requiring plating dishes with a gelatin coat and then culturing for an additional 72 hours. There has been significant progress in the past 3 – 4 years on determining embryo quality in an IVF cycle and some of the same technology should be applied to quality control testing.

    • Micah Hill

      I completely agree. Very nice study!

  • Micah Hill

    Congratulations Ravi on a very nice study. I like your endpoint being further downstream than blastocyst formation and your dose-response figures were convincing. How does ICM outgrowth assay compare with the other 2 assays from the standpoint of cost and time dedication?

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