pH stability of human preimplantation embryo culture media: effects of culture and batches

Abstract

Research question: How stable is the pH of human preimplantation embryo culture media during IVF culture and is there variation in pH between batches of culture media?

Design: To evaluate pH stability, three batches of three culture media were incubated in triplicate without embryos (sham culture) at CO₂ levels recommended by the manufacturers (5% or 6%) for 4 days. To evaluate differences in pH between batches, the pH of three batches of five culture media was measured in triplicate during 1 day of sham culture. Linear mixed models were used for the analysis.

Results: An increase in pH during 4 days of culture was found in all three culture media, but the observed increased values were within the generally accepted range for clinical practice (pH 7.2-7.4). One medium was pH 7.1 in the first 2 days, but this was within the range provided by the manufacturer for that medium. Three out of five analysed media showed batch variation in pH that exceeded the generally accepted range for clinical practice.

Conclusions: A relevant difference in pH was found between batches of human preimplantation embryo culture media. This suggests that the CO₂ level of incubators may need to be adjusted for new batches of culture medium based on measured pH, to anticipate batch variability and safely accommodate limited pH increase over time. This study was unable to identify the cause of the differences in pH between batches, and further investigation on a larger number of batches and other media seems warranted.

Keywords: Culture medium; Human preimplantation embryo culture; IVF; Intracytoplasmic sperm injection; pH.