Mitochondrial DNA in Fresh Versus Frozen Embryo Culture Media of PCOS Women Undergoing In Vitro Fertilization
Condition: Polycystic Ovary Syndrome; Assisted Reproduction
MeSH terms: Polycystic Ovary Syndrome; Syndrome
Study type: Observational

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)
    

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).
      
Biospecimen description: 
      embryo culture media from day 3 morphologically good embryos will be collected after embryo
      transfer of fresh and thawed cycles. The media will be immediately frozen into sterile, PCR
      clean (free from DNA, DNase, RNase and PCR inhibitors) tubes and stored at - 20 C until
      nucleic acid purification