Validation of Addition of Uterine Fluid to Human Embryo Culture Medium

Infertility Clinical Trial

Official title:

Validation of Addition of Uterine Fluid to Human Embryo Culture Medium

Clinical Trial Details — Status: Recruiting

Administrative data

• NCT number: NCT03436758
• Other study ID #: 1607-MUR-055-JL
• Status: Recruiting
• Phase: N/A
• First received: February 12, 2018
• Last updated: February 12, 2018
• Start date: January 30, 2018
• Est. completion date: December 31, 2019

Study information

• Verified date: February 2018
• Source: IVI Murcia
• Contact: Juan C Martinez-Soto, PhD
• Phone: +34968200031
• Email: juancarlos.martinez[@]ivi.es
• Is FDA regulated: No
• Study type: Observational

Clinical Trial Summary

Infertility or infertility affects about 15% of couples of reproductive age. It is estimated that 80 million people around the world suffer from this problem. Assisted reproduction techniques (ART) use culture media (during in vitro fertilization or early embryo development) with protein sources that are very different from natural sources. This media could produce an added stress to the gametes and embryos that could cause epigenetic alterations and health effects during adult life.

Our working hypothesis is based on studies in animal models (pig and cow), in which it was observed that the culture media with reproductive fluids used as additives instead of conventional sources of proteins (such as serum albumin) , produce embryos with an epigenetic profile closer to that of embryos generated in the maternal oviduct. Moreover, with these fluids, blastocysts obtained have a greater number of cells and hatchability than those produced with serum albumin alone.

Therefore, the University of Murcia, with an extensive experience in this area, and the IVI Murcia (Valencian Infertility Institute) research team have come together to launch this research project in order to determine the advantages of the use of human reproductive fluids as additives in embryonic culture media. To do this, 2 specific objectives are proposed,:

1. Creation of the first collection of human uterine fluid samples for Assisted Reproduction use.

2. To evaluate the use of uterine fluid as a media supplement in the culture media for assisted reproduction techniques, by evaluating embryo quality cultured with autologous fluid from voluntary patients (autologous culture)

This achievement would allow us the development of protocols in the nearest ART physiological conditions which represent not only a technical challenge but a biomedical responsibility that must be addressed to prevent future diseases of the offspring.

Description:

1. Creation of the first collection of human uterine fluid samples for Assisted Reproduction use. Female reproductive system will be collected at the Virgen de la Arrixaca University Clinical Hospital from surgical pieces of patients undergoing surgical procedures. IVI Murcia Clinics uterine fluids from oocyte donors will be collected. These fluids will be store in the Arrixaca Hospital Biobank for future researches.

2. Validation of the addition of uterine fluid to human embryonic culture media:

Couples undergoing assited Reproduction Treatment in IVI Murcia will be asked for their inclusion in this study. Uterine fluid will be take 48 hours after peak of LH (luteinizing hormone), in natural cycle. Oocytes from these patients, will be divided into two groups: Half of the zygotes of each patient, control group, will be grown in the conditions usually used in the clinic, and the other half, experimental group, will add 1-5% of uterine fluid from the patient (v / v)

On day 3 and on day 5 of culture the quality of the embryos in both groups will be assessed according to the usual criteria of the clinic and the best quality and highest probability of implantation will be transferred, following the usual clinical practice. The samples of uterine fluid, after the initial processing in the IVI-Murcia clinic will be transferred to the Department of Physiology of the University of Murcia where they will be fractionated to perform a quality control according to the following specifications:

Uterine fluid pH 7.0-7.6 Osmolarity (mOsm / kg) 260-320 Endotoxin (EU / mL) <0.15 Sterility No growth

Only samples that meet these criteria will continue to be part of the study

Recruitment information / eligibility

• Status: Recruiting
• Enrollment: 27
• Est. completion date: December 31, 2019
• Est. primary completion date: October 30, 2019
• Gender: All
• Age group: 18 Years to 39 Years

Eligibility

Inclusion Criteria:

• Couples in icsi treatment

Woman:

BMI between 17 and 30.Under controlled ovarian stimulation and at least 8 metaphase II oocytes collected

Man:

Sperm concentration higher than 5 million sperm / ml

Exclusion Criteria:

• Woman:

Diagnosis of endometriosis Uterine pathology that compromises embryonic development Patients undergoing treatment for repeat abortion or implantation failures Abnormal karyotype Patients included in the Pre-Implantation Genetic Diagnosis (DPI) program

Male:

Men whit abnormal karyotype, High values of DNA fragmentation Abnormal FISH (Fluorescence in situ Hybridization) Sperm from epididymal aspirate or testicular biopsy

Related Conditions & MeSH terms

• Infertility

Locations

Country	Name	City	State
Spain	IVI Murcia	Murcia

Sponsors (2)

Lead Sponsor	Collaborator
IVI Murcia	Universidad de Murcia

Country where clinical trial is conducted

Spain, 

References & Publications (7)

Avilés M, Gutiérrez-Adán A, Coy P. Oviductal secretions: will they be key factors for the future ARTs? Mol Hum Reprod. 2010 Dec;16(12):896-906. doi: 10.1093/molehr/gaq056. Epub 2010 Jun 28. Review. — View Citation

Belva F, Bonduelle M, Roelants M, Michielsen D, Van Steirteghem A, Verheyen G, Tournaye H. Semen quality of young adult ICSI offspring: the first results. Hum Reprod. 2016 Dec;31(12):2811-2820. Epub 2016 Oct 5. — View Citation

Chiu PC, Chung MK, Koistinen R, Koistinen H, Seppala M, Ho PC, Ng EH, Lee KF, Yeung WS. Cumulus oophorus-associated glycodelin-C displaces sperm-bound glycodelin-A and -F and stimulates spermatozoa-zona pellucida binding. J Biol Chem. 2007 Feb 23;282(8):5 — View Citation

Coy P, Cánovas S, Mondéjar I, Saavedra MD, Romar R, Grullón L, Matás C, Avilés M. Oviduct-specific glycoprotein and heparin modulate sperm-zona pellucida interaction during fertilization and contribute to the control of polyspermy. Proc Natl Acad Sci U S — View Citation

Coy P, Grullon L, Canovas S, Romar R, Matas C, Aviles M. Hardening of the zona pellucida of unfertilized eggs can reduce polyspermic fertilization in the pig and cow. Reproduction. 2008 Jan;135(1):19-27. — View Citation

Coy P, Jiménez-Movilla M, García-Vázquez FA, Mondéjar I, Grullón L, Romar R. Oocytes use the plasminogen-plasmin system to remove supernumerary spermatozoa. Hum Reprod. 2012 Jul;27(7):1985-93. doi: 10.1093/humrep/des146. Epub 2012 May 3. — View Citation

Gabler C, Chapman DA, Killian GJ. Expression and presence of osteopontin and integrins in the bovine oviduct during the oestrous cycle. Reproduction. 2003 Dec;126(6):721-9. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	embryo quality	Number of embryo with A or B quality according to ASEBIR (Spanish association for the study of the biology of reproduction) criteria	embryo quality will be analyze on day 3 and 5 after fertilization
Secondary	Pregnancy rate	serum levels of chorionic gonadotropin Beta (Beta-hCG)	between 14-16 days after fertilization
Secondary	ongoing pregnancy	ultrasonography	16-18 week after fertilization
Secondary	implantation rate	number of embryos implanted/number of embryos transferred x100	24 days after fertilization
Secondary	pregnancy loss rate	number of miscarriages/ number of ongoing pregnancy x100	10 days after B-hCG evaluation at end of pregnancy