hCG-activated PBMC-therapy in RIF Patients

248 women with the history of implantation failure volunteered to receive PBMC-therapy. After immunologic consultation and doing flow cytometry analysis, 100 women with at least three IVF/ET failure who had low Th-17/Treg ratio in comparison with healthy control were enrolled in this study. These 100 patients divided randomly into two groups, 50 patients received PBMC and 50 patients as the control group received PBS. PBMCs were obtained from patients themselves five days before embryo transfer (ET) and were cultured with hCG for 48 hours. Frothy-eight hours later, PBMCs were then administered into the uterine cavity of that patient from the study group two days before ET. PBS was inseminated into the uterine cavity of the control group instead of PBMC. The concentration of inflammatory cytokines was examined in the supernatant of cultured PBMCs 2, 24 and 48 hour after incubation by ELISA. The pregnancy occurrence was confirmed 12 days after ET through positive pregnancy test (β-hCG test). The success of implantation and the occurrence of clinical pregnancy were evaluated by ultrasound through the observation of the number and the location of gestational sacs at 5-6 weeks and confirming the embryo heart pulsation.

• Peripheral Blood Monouclear Cell (Other), also known as PBMC: Blood samples (20 mL) were taken from individual patients at the time of ovulation induction. Then PBMC were isolated and cultured (20-30 million cells) 48 hour at the presence of hCG (10IU/ml daily). Afterward,15-20 million PBMCs in 500 microlitres PBS were injected into the uterine cavity two days before embryo transfer (ET) using ET catheter.
• Phosphate Baffer Saline (Other), also known as PBS: Only 500 microlitres PBS will be injected into the uterine cavity, instead of PBMCs, two days before embryo transfer (ET) using ET catheter.

Primary outcomes

• pregnancy occurrence ["12 days after embryo transfer"]: Laboratory tests (beta-hCG test)
• examination of inflammatory cytokines (IL-1B, TNF-a and INF-Y) secretion levels in supernatant ["48 hours after culturing"]: ELISA technique

Secondary outcomes

• Clinical pregnancy rate in patients with recurrent implantation failure (RIF) ["in 5-6 weeks"]: detection the number and location of gestational sacs by ultrasound
• The live birth rate by patients with recurrent implantation failure (RIF) ["After about 9 months of positive βHCG test"]: By gynecologists and obstetricians will monitor
• The miscarriage rate in patients with recurrent implantation failure (RIF) ["When ever during the pregnancy period (up to 9 months)"]: By gynecologists and obstetricians will monitor

Sex: Female, ages: 20 Years to 45 Years, accepts healthy volunteers

All patients were evaluated in accordance with the following inclusion and exclusion criteria;

Inclusion Criteria:

1. Having at least three implantation failures following IVF
2. Having primary infertility
3. Age under 45 years old
4. Having regular menstrual cycles
5. BMI under 30

Exclusion Criteria:

1. Having polycystic ovary syndrome
2. The presence of uterine pathology;
3. Poor ovarian reserve
4. Having chromosomal abnormalities
5. Presence of auto anti-bodies such as anti-TPO, anti-TG, ACA, APA, ANA, and anti-dsDNA
6. Presence of mutations involving the coagulation system such as deficiency of factor XII, Pro C, Pro S
7. Positive HIV, HCV or HBV tests

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• Mohammad Nouri, Ph.D, Study chair, Head of SCARM institute
• Mehdi Yousefi, Ph.D, Study director, SCARM institute