Clinical trial NCT03820167: Mitochondrial DNA in Fresh Versus Frozen Embryo Culture Media of PCOS Women Undergoing In Vitro Fertilization
Official title: Mitochondrial DNA in Fresh Versus Frozen Embryo Culture Media of Polycystic Ovarian Syndrome Patients Undergoing In Vitro Fertilization
Study type: OBSERVATIONAL
Overall status: COMPLETED
Conditions: Polycystic Ovary Syndrome
Assisted Reproduction
Lead sponsor: Cairo University
Sponsor class: OTHER
First posted: 2019-01-29
Enrollment: 90
Sex: FEMALE
Minimum age (years): 20
Maximum age (years): 38
Standard ages: ADULT
Accepts healthy volunteers: no
Observational model: CASE_CONTROL
Time perspective: PROSPECTIVE
Sampling method: PROBABILITY_SAMPLE
Study population: Detailed history taking with special emphasis on:

1. Personal history (name, age, marital state, parity, address).
2. Menstrual history (duration, cycle, amount, regularity, last menstrual period).
3. Medical history for acne and hirsutism.

Clinical examination with special emphasis on:

1. Height and weight of the patients to calculate the BMI (weight in kg/ height in m2).
2. Examinationfor presence of acne and/ or hirsutism (patients with score \> 8 on Ferryman \& Gallawey score were considered as hirsute).
3. Ultrasound examination to confirm diagnosis of PCOS (Using the vaginal probe).

Brief summary: Ninety women with PCOS candidate for ICSI were randomized to 2 groups:Group A: fresh samples of PCOS patients undergoing IVF (no= 45) and Group B: frozen samples of PCOS patients undergoing IVF (no= 45). Morphologically good embryos will be cultured in a culture dish and the supernatants will be collected freshly from culture system at day 3 and stored at -20 ̊ c until being tested. Morphologically good embryos scheduled for freezing will be cryopreserved for less than one year and thawed, the supernatant will be collected and stored at -20 ̊ c until being tested. Quantification of mtDNA in fresh and frozen culture media using qPCR technique.The template DNA prepared and the dilution standards prepared will be used for qPCR to determine the amount of MtDNA and nuclear DNA in the sample using QuantiTect SYBR Green PCR Kit

Detailed description: Ninety women with PCOS candidate for ICSI were randomized to 2 groups:Group A: fresh samples of PCOS patients undergoing IVF (no= 45) and Group B: frozen samples of PCOS patients undergoing IVF (no= 45). Morphologically good embryos will be cultured in a culture dish and the supernatants will be collected freshly from culture system at day 3 and stored at -20 ̊ c until being tested. Morphologically good embryos scheduled for freezing will be cryopreserved for less than one year and thawed, the supernatant will be collected and stored at -20 ̊ c until being tested. Quantification of mtDNA in fresh and frozen culture media using qPCR technique.The template DNA prepared and the dilution standards prepared will be used for qPCR to determine the amount of MtDNA and nuclear DNA in the sample using QuantiTect SYBR Green PCR Kit (Qiagen)

Eligibility criteria: Inclusion Criteria:

1. Age: 20-35 yrs.
2. BMI: ≤25.
3. PCO: diagnosed by Rotterdam criteria (at least two of the following):

   1. Polycystic ovary morphology with one ovary being sufficient for diagnosis.
   2. Hyperandrogenism: clinically or biochemically.
   3. Anovulation or oligo-ovulation.
4. Gravidity and parity: Nulligravida.
5. Compliance: primary infertility

Exclusion Criteria:

* Known medical problems as diabetes mellitus, hypertension, cardiac, hepatic or renal problem.
* Special habits of medical importance as smoking.
* Other endocrinopathies

Primary outcomes:
measure: clinical pregnancy rate
timeFrame: 4 weeks after embryo transfer
Secondary outcomes:
Other outcomes:
Locations:
city: Giza
country: Egypt
facility: Ganna IVF center
state: Cairo
Study officials:
affiliation: Professor
name: Ahmed Maged, MD
role: PRINCIPAL_INVESTIGATOR
References: