1. Peripheral blood mononuclear cells composition for
the autoimmune infertility treatment in IVF practice
A.M. Feskov, I.A. Feskova, I.S. Zhylkova, N.N. Sotnik, V.A. Feskov
Clinic of Professor Feskov A.M., Ukraine
and
S. V. Zhilkov
MEZADATA Medical LLC, USA
Presented in Prague, Czech Republic, December 2014

2. Embryo implantation:
who is responsible?
1. Embryo quality
VS
2. Endometrium quality

3. Scheme of the best result:

4. Role of immune system
in embryo implantation
• Maternal immune system recognizes certain presence of the developing
and implanting embryo in the Fallopian tube and uterus cavity due to
embryo‐ and species‐specific signals such as degraded products of zona
pellucida glycoprotein and/or HCG.
• Effector immune cells move to the ovary and the endometrium via blood
circulation to regulate corpus luteum (CL) function and induce
endometrial differentiation.
• Local immune cells at the implantation site also contribute to induction
of embryo invasion, secreting chemoattractants by HCG stimulation.
• To achieve smooth transformation into CL of pregnancy, it is reasonable
to expect that immune cells transmit information from the developing
embryo to CL before embryo implantation. In support of this hypothesis,
the HLA‐DR and leukocyte functional antigen (LFA)‐3/CD58, which could
mediate the direct interaction with T lymphocytes, were expressed on
human CL during corpus luteum formation.

5. Immune system and peripheral blood
mononuclear cells: what has been
already known?
Feskov, A., et al. US 61/629,651, 11/23/2011
TITLE OF THE INVENTION: METHOD OF IN VITRO FERTILIZATION WITH DELAY OF EMBRYO
TRANSFER AND USE OF PERIPHERAL BLOOD MONONUCLEAR CELLS
… The in vitro fertilization method of the invention provides the use of the PBMCs obtained by the above culturing
technique in order to improve pregnancy inception…
Okitsu O, Kiyokawa M, Oda T, Miyake K, Sato Y, Fujiwara H. J Reprod Immunol. 2011 Dec;92(1-
2):82-7. doi: 10.1016/j.jri.2011.07.001. Epub 2011 Oct 27
Intrauterine administration of autologous peripheral blood mononuclear cells increases clinical
pregnancy rates in frozen/thawed embryo transfer cycles of patients with repeated implantation
failure
…intrauterine administration of autologous PBMC freshly isolated from patients, effectively improves embryo implantation
in patients with three or more IVF failures…
Feskov O.M., Feskova I.A., Zhylkova I.S., Bezpechna I.M., Blazhko O.V. World of Medicine and
Biology, №3, pp. 132-134, 2011.
The influence of intrauterine administration of peripheral blood mononuclear cells on
implantation rates in “fresh” and “cryo” IVF programs
…This study showed that intrauterine application of PBMC definitely (р <0,05) increased the implantation rates and
clinical pregnancy rates both after embryo transfer in fresh IVF cycles and after the thawed embryos transfer…
Yu N1, Yang J, Guo Y, Fang J, Yin T, Luo J, Li X, Li W, Zhao Q, Zou Y, Xu W. Am J Reprod Immunol. 2014
Jan;71(1):24-33. doi: 10.1111/aji.12150. Epub 2013 Aug 1.
Intrauterine administration of peripheral blood mononuclear cells (PBMCs) improves endometrial receptivity in
mice with embryonic implantation dysfunction
…Embryo implantation dysfunction model group showed a significant decrease in pregnancy rate, implantation sites and
the expression of both the endometrial LIF and VEGF during the implantation window. EID with PBMCs group showed a
higher pregnancy rate and endometrial LIF and VEGF expression compared to EID group…

6. Aim of present
investigation:
Study question: The influence of the intrauterine peripheral
blood mononuclear cells (PBMCs) application on the embryo
implantation rates was studied in patients with repeated
implantation failures (RIF) in IVF cycles.
Study design: The study population consists of 180 infertile
female patients, who already had two or more unsuccessful IVF
attempts with transfer of the high quality embryos in the past.

7. Participants:
Group 1:
90 patients with the median age of 34.2±4.3 years old.
Intrauterine PBMCs were applied in group 1 consequentially:
1) First in “fresh” IVF cycle: PBMCs were applied for 90
patients. 28 clinical pregnancies were achieved after 90 ET
of “fresh” embryos. The rest 62 patients moved to the cryo‐
IVF attempt.
2) For these 62 patients, PBMCs were again applied prior
transfer of the vitrified embryos that were under cryo‐
preservation after first IVF attempt above. 26 clinical
pregnancies were achieved after 62 ET of thawed embryos.

8. Participants:
Group 2:
90 patients with the median age of 35.2±5.6 years old.
Intrauterine PBMCs were not applied in group 2:
1) First in “fresh” IVF cycle 20 clinical pregnancies were
achieved after 90 ET of “fresh” embryos. The rest 70
patients moved to the cryo‐IVF attempt.
2) For these 70 patients, the transfer of the vitrified embryos
was done. 15 clinical pregnancies were achieved after 70 ET
of thawed embryos.

9. IVF procedure
COS (controlled ovary stimulation): protocol with
a‐GnRH was used for patients of both groups.
The period of ovary stimulation for every patient
was no less then 10 days. To the moment of
transvaginal puncture, the average size of
follicles was about 18 mm. To maintain the luteal
phase, the medicines with progesterone were
used for patients in both groups.
Embryo culture: After oocytes were retrieved,
they were cultured in Universal IVF Medium
(Medicult) in 6.0 % CO2 and 37.0ºC. The embryos
were cultured in Global Total Medium (LifeGlobal
Group).

10. Preparation and intrauterine
application of PBMC
1. PBMCs were isolated by Ficoll‐Hypaque centrifugation.
2. After extraction, PBMCs were cultured in vitro. The conditions of culture were
carbon dioxide atmosphere and 37 ºC.
3. Fresh PBMCs were obtained from the same patient few days after the first
portion of PBMCs culture. Freshly isolated and cultured PBMCs were mixed and
transferred to uterus cavity.

11. Results:
Clinical Pregnancy Rate
Group 1 (with PBMCs) Group 2 (without PBMCs)
Fresh‐IVF
31.1%
(28 pregnancies after 90 ET)
22.2%
(20 pregnancies after 90 ET)
Cryo‐ IVF
41.9%
(26 pregnancies after 62 ET)
21.4%
(15 pregnancies after 70 ET)
Total
60.0%
(54 pregnancies for 90 patients)
38.9%
(35 pregnancies for 90 patients)

12. Results:
Clinical Pregnancy Rate
Group 1 (with PBMCs) Group 2 (without PBMCs)
Cryo‐ IVF
41.9%
(26 pregnancies after 62 ET)
21.4%
(15 pregnancies after 70 ET)
Special attention should be addressed to the clinical pregnancy rate after the
transfer of thawed embryos in group with PBMCs application. It is further
demonstrated that the combining of delayed or Cryo-IVF technique (cryopreservation
IVF technique or “Cryo-IVF”) with enhanced preparation of the woman’s uterus by the
administration of her own PBMCs prior embryo transfer does result in a significant
increase in the rate of pregnancy of infertile women as compared to previously-known
IVF procedures; possibly it is a synergistic effect both of allowing the female patient’s
autoimmune system to regain hormonal balance after COS and allowing the inception
process to be realized in uterine cavity of improved quality.

13. Conclusions:
• Intrauterine application of PMBC increases rates of
the embryo implantation and can be used for
patients with repeated implantation failures (RIF).
• For patients with RIF, the clinical pregnancy rate
after applying of the PBMC is significantly higher
compare with patients without PBMC use (p<0.05);
this is observed for embryo transfers both in the
“fresh” and “cryo” IVF cycles.
• PBMC method might be also effective in infertility
treatment of older female patients, because among
this age group the RIF patients constitute significant
majority.

14. Thank you for your attention !
Contact: Zhilkova@Feskov.com.ua