Optimizing the Temperature for Embryo Culture in In Vitro Fertilization

Optimizing the Temperature for Embryo Culture in In Vitro Fertilization: A Randomized Controlled Trial

Many human and animal studies over the past 40 years have revealed that ovaries and Fallopian tubes have a lower temperature than core body temperature, especially around the time of ovulation and early embryo development. Whether average core body temperature, 37 degrees Celsius, is ideal temperature for culture has not been prospectively evaluated. The purpose of this study is to determine whether lowering the incubator temperature by 1 degree Celsius will result in meaningful improvements in embryo development and pregnancy rates.

Patients participating in this study will undergo routine in vitro fertilization (IVF) stimulation. Following egg retrieval, patients with at least 8 mature eggs will have those eggs divided randomly into two groups. Both groups will then under ICSI (intra-cytoplasmic sperm injection) to fertilize the eggs. The groups will randomly be assigned to control or study condition. The control condition group will be cultured at 37 degrees Celsius, the current standard of care. The study group will be cultured at 36 degrees Celsius. All other culture conditions will be kept identical and embryos will be assessed on day 3 and day 5 of development.

On day 5, all well-developed blastocysts will undergo biopsy for Comprehensive Chromosomal Screening (CCS), which will evaluate the chromosomal normalcy of the embryo . Patients will undergo transfer on the morning of day 6 and the highest quality, chromosomally normal embryo from each group (the 36 degree cultured embryos and the 37 degree cultured embryos) will be transferred. If only 1 embryo is available for transfer, the patient will have a single embryo transfer.

All embryos available for cryopreservation will undergo biopsy for CCS before being frozen. Some arrested embryos from each temperature group will also undergo genetic analysis to evaluate chromosomal normalcy or mosaicism.

The female patient will undergo a blood draw at 9 weeks gestation and buccal swabs will be obtained from the infant(s) after birth.

Study Overview

• Status: Completed
• Conditions: Infertility
• Intervention / Treatment: Other: 36 degrees Celsius

• Study Type: Interventional
• Enrollment (Actual): 55
• Phase: Not Applicable

Contacts and Locations

Study Locations

• United States
  • New Jersey
    • Morristown, New Jersey, United States, 07960
      • Reproductive Medicine Associates of new Jersey
  • Pennsylvania
    • Allentown, Pennsylvania, United States, 18104
      • Reproductive Medicine Associates of Pennsylvannia

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 41 years (Adult)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: Female

Description

Inclusion Criteria:

• No more than 1 prior failed cycle at RMA NJ facility; up to 2 prior failed cycles at another facility pending investigator review
• Female partner between 18 and 41 at onset of IVF cycle
• Normal day 3 FSH (< or = 12) in RMA NJ laboratory
• Basal antral follicle count of at least 10
• Male partner with greater than 100,000 total motile spermatozoa per ejaculate (donor sperm is ok)
• BMI < or = 32 kg/m2

Exclusion Criteria:

• Diagnosis of chronic anovulation (cycles occuring typically less often than every 90 days)
• Use of testicular aspiration or biopsy procedures to obtain sperm
• Unevaluated ovarian mass or diagnosis of surgically-confirmed stage IV endometriosis
• Presence of hydrosalpinges that communicate with the uterine cavity
• History of endometrial insufficiency
• Any contraindication to undergoing IVF or gonadotropin stimulation
• Prior poor fertilization rate- less than 50% of mature eggs
• Prior poor blastulation rate- less than 10% of 2PNs (day 1 embryos)

Study Plan

How is the study designed?

Design Details

• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: 36 degree group; 36 degrees Celsius for the incubators is the experimental condition in this study.	Other: 36 degrees Celsius; Embryos in this group will be cultured in incubators set to 36 degrees Celsius.; Other Names: IVF, PGD, embryos, culture, CCS, infertility
No Intervention: 37 degree group; 37 degrees Celsius is the standard incubator temperature for the culture of embryos.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Useable Blastocyst Formation Rate	Proportion of mature eggs developing into expanded blastocysts (Gardner grade 3-6) of sufficient quality to transfer or cryopreserve will be documented for each group (36 degree and 37 degree group).	approximately 1 week

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Fertilization rate	The proportion of mature eggs that underwent ICSI and are normally fertlized (documented as forming 2 pro-nuclei (2PNs)) on the day after egg retrieval will be documented for each group (36 degree and 37 degree groups)	approximately 2 days
Cleavage Rate	Proportion of properly fertlized embryo (2PNs on day 1) that undergo further celluar division will be documented for each group (36 degree and 37 degree group).	approximately 1 week
Blastocyst Formation Rate	Proportion of mature oocytes that go on to develop into blastocysts in each group (36 degree group and 37 degree group) will be documented.	approximately 1 week
Implantation Rate	Proportion of transferred embryos resulting in a clinically documented pregnancy will be documented for each group (36 degree and 37 degree group).	approximately 2 weeks
Aneuploidy Rate	Rate of chromosomal abnormalities among the embryos in each group (36 degree and 37 degree groups) as determined by polymerase chain reaction (PCR) based comprehensive chromosome screening (CCS).	approximately 1 month
Mosaicism Rate	Proportion of embryos that arrest at the cleavage stage which show evidence of more than one chromosomal lineages (mosaicism) when analyzed my microarray documented in each group (36 degree and 37 degree group).	approximately 1 month

Collaborators and Investigators

• Sponsor: Reproductive Medicine Associates of New Jersey
• Investigators: Principal Investigator: Richard T Scott, MD, Reproductive Medicine Associates of new Jersey

Publications and helpful links

General Publications

• Hong KH, Lee H, Forman EJ, Upham KM, Scott RT Jr. Examining the temperature of embryo culture in in vitro fertilization: a randomized controlled trial comparing traditional core temperature (37 degrees C) to a more physiologic, cooler temperature (36 degrees C). Fertil Steril. 2014 Sep;102(3):767-73. doi: 10.1016/j.fertnstert.2014.06.009. Epub 2014 Jul 17.

Helpful Links

• Reproductive Medicine Associates of New Jersey

Study record dates

Study Major Dates

• Study Start: January 1, 2012
• Primary Completion (Actual): January 1, 2014
• Study Completion (Actual): July 1, 2014

Study Registration Dates

• First Submitted: January 3, 2012
• First Submitted That Met QC Criteria: January 6, 2012
• First Posted (Estimate): January 9, 2012

Study Record Updates

• Last Update Posted (Estimate): March 26, 2015
• Last Update Submitted That Met QC Criteria: March 25, 2015
• Last Verified: March 1, 2015

More Information

Terms related to this study

• Keywords: IVF; infertility; temperature; PGD; CCS; culture; embryo; incubator; incubator-36 degrees Celsius; incubator-37 degrees Celsius
• Additional Relevant MeSH Terms: Infertility
• Other Study ID Numbers: RMA-2011-02