Composition of commercial media used for human embryo culture

Capsule:
There is considerable biologically relevant variation in the compositions of commercially available culture media. This information provides a framework for future analysis of long-term effects of in vitro culture and optimization of culture media.

Authors:
Dean E. Morbeck, Ph.D., Rebecca L. Krisher, Ph.D., Jason R. Herrick, Ph.D., Nikola A. Baumann, Ph.D., Thomas Moyer, Ph.D., Dietrich Matern, M.D., Ph.D.

Volume 102, Issue 3, Pages 759-766

Abstract:

Objective:
To determine the composition of commercially available culture media and test whether differences in composition are biologically relevant in a murine model.

Design:
Experimental laboratory study.

Setting:
University-based laboratory.

Animal(s):
Cryopreserved hybrid mouse one-cell embryos were used in experiments.

Intervention(s):
Amino acid, organic acid, ions, and metal content were determined for two different lots of media from Cook, In Vitro Care, Origio, Sage, Vitrolife, Irvine CSC, and Global. To determine whether differences in the composition of these media are biologically relevant, mouse one-cell embryos were thawed and cultured for 120 hours in each culture media at 5% and 20% oxygen in the presence or absence of protein in an EmbryoScope time-lapse incubator.

Main Outcome Measure(s):
The compositions of seven culture media were analyzed for concentrations of 39 individual amino acids, organic acids, ions, and elements. Blastocyst rates and cell cycle timings were calculated at 96 hours of culture, and the experiments were repeated in triplicate.

Result(s):
Of the 39 analytes, concentrations of glucose, lactate, pyruvate, amino acids, phosphate, calcium, and magnesium were present in variable concentrations, likely reflecting differences in the interpretation of animal studies. Essential trace elements, such as copper and zinc, were not detected. Mouse embryos failed to develop in one culture medium and were differentially affected by oxygen in two other media.

Conclusion(s):
Culture media composition varies widely, with differences in pyruvate, lactate, and amino acids especially notable. Blastocyst development was culture media dependent and showed an interaction with oxygen concentration and presence of protein.

  • Juan Manuel Mormoy

    Hi. Nice work.

    I just was wondering if in the supplementary table 5, the “cc2” parameter is ok. Since your values are aproximately 20 hours while papers published has shown values of 11-12 hours. Thank you so much.
    Sincerely
    Dr. Moreno-Moya

    • Dean Morbeck

      Dear Dr. Moreno-Moya,
      You are correct that human embryos remain at the 2-cell stage for 10-2 hours. This work was with mouse embryos, which have a very long 2-cell stage that typically lasts >20 hours. We are not sure why there is such a difference between the species, but it is quite possible that the high rate of early genome activation present in murine embryos at the 2-cell stage is involved.
      Dean
      Dean

      • Juan Manuel Mormoy

        Wow thats awesome! Thank you so much for your fast reply Dean.

        Sincerely
        Dr Moreno-Moya

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