Relationship of reactive oxygen species levels in day 3 culture media to the outcome of in vitro fertilization/intracytoplasmic sperm injection cycles

Received 2 August 2009; received in revised form 3 December 2009; accepted 8 December 2009. published online 05 February 2010.
Corrected Proof

Objective

To examine the relationship of early human embryonic development parameters to day 3 reactive oxygen species (D-3 ROS) levels in culture media.

Design

Prospective study.

Setting

Tertiary care hospital.

Patient(s)

Patients were undergoing IVF (n = 92; 36 with intracytoplasmic sperm injection [ICSI]).

Intervention(s)

The D-3 ROS levels in sample and control of each embryo culture dish were measured by the chemiluminescence method using a luminol probe.

Main Outcome Measure(s)

Embryo quality (days 3 and 5) and pregnancy rates (PR).

Result(s)

The D-3 ROS level was significantly lower in pregnant cycles 26.8 ± 13.9 × 106 cpm (counted photon per minute) versus nonpregnant cycles 66.4 ± 39.4 × 106 cpm. This relationship was maintained when the cycles were stratified to conventional IVF (27.1 ± 14.95 vs. 67.0 ± 39.9 × 106 cpm) or ICSI (25.6 ± 12.75 vs. 65.5 ± 39.7 × 106 cpm). After controlling for all variables, D-3 ROS levels were negatively correlated with blastocyst development rate as well as PR. Odds ratio (OR) (95% confidence interval [CI]) of clinical pregnancy corresponding to a 10 × 106 cpm increase in D-3 ROS was 0.47 (0.30–0.74) for ICSI and 0.56 (0.37–0.85) for IVF.

Conclusion(s)

During extended in vitro culture, ROS generated in culture media by day 3 may be an important biochemical marker for blastulation. An increase of 10 units in D-3 ROS may decrease the clinical pregnancy by 41%.

Key Words: Reactive oxygen species, fertilization, blastulation, ICSI, IVF, extended embryo culture

a Department of Obstetrics and Gynecology, University Hospitals Case Medical Center, Case Western Reserve University, Cleveland, Ohio

b Center for Reproductive Medicine, Glickman Urological and Kidney Institute, & Ob-Gyn and Women's Health Institute, Cleveland Clinic, Cleveland, Ohio

Reprint requests: Ashok Agarwal, Ph.D., Director, Center for Reproductive Medicine, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195 (FAX: 216-445-6049).

M.A.B. has nothing to disclose. R.Z.M. has nothing to disclose. J.M.G. has nothing to disclose. R.S. has nothing to disclose. T.F. has nothing to disclose. M.F.A.H. has nothing to disclose. A.A. has nothing to disclose.

Presented in part at the 63rd Annual Meeting of the American Society for Reproductive Medicine, October 13–17, 2007, Washington, DC.

PII: S0015-0282(09)04206-X

doi:10.1016/j.fertnstert.2009.12.020