Attempts to improve human ovarian transplantation outcomes of needle-immersed vitrification and slow-freezing by host and graft treatments.
To investigate if needle-immersed vitrification or slow-freezing yields better implantation results for human ovarian tissue and which method benefits more when combined with the "improvement protocol" of host melatonin treatment and graft incubation with biological glue + vitamin E + vascular endothelial growth factor-A.



Human ovarian tissue was preserved by needle-immersed vitrification or slow-freezing and transplanted into immunodeficient mice, either untreated (groups A and C, respectively) or treated with the improvement protocol (groups B and D, respectively). Grafted and ungrafted slices were evaluated by follicle counts, apoptosis assay and immunohistochemistry for Ki67 and platelet endothelial cell adhesion molecule (PECAM).



Follicle number in the recovered grafts was limited. The number of atretic follicles was significantly higher after vitrification with/without the improvement protocol and slow-freezing than that after slow-freezing + the improvement protocol. Stroma cell apoptosis was the lowest in the group D. PECAM staining showed a peripheral and diffuse pattern in the group D (mostly normal follicular morphology) and a diffuse pattern in all other groups (few follicles, mostly atretic), with significantly higher diffuse levels in the vitrification groups. Ki67 staining was identified in all normal follicles. Follicles did not survive transplantation in the vitrification groups.



Ovarian sample preparation with slow-freezing + the improvement protocol appears to yield better implantation outcomes than needle-immersed vitrification with/without the improvement protocol. The real quality of frozen tissue can be assessed only after grafting and not after thawing/warming.
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel. ronita@clalit.org.il.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel. ronita@clalit.org.il.
Ronit
R
Abir
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Benjamin
B
Fisch
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Noa
N
Fisher
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.|In Vitro Fertilization Unit, Lis Maternity and Women's Hospital, Tel Aviv Sourasky Medical Center, 6423906, Tel Aviv, Israel.
Nivin
N
Samara
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Galit
G
Lerer-Serfaty
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Faculty of Health Sciences, Goldman Medical School, Ben Gurion University of the Negev, Beer Sheva, 8410501, Israel.
Roei
R
Magen
Department of Nephrology, Rabin Medical Center, Felsenstein Research Center 49100 and Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Michal
M
Herman-Edelstein
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Avi
A
Ben-Haroush
Infertility and IVF Unit, Beilinson Women Hospital, Rabin Medical Center, Petach Tikva, 49100, Israel.|Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Anat
A
Stein
Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.|Infertility and IVF Unit, Department of Obstetrics and Gynecology, The Chaim Sheba Medical Center, Ramat Gan, 52621, Israel.
Raoul
R
Orvieto
D000328:Adult; D000818:Animals; D017209:Apoptosis; D002470:Cell Survival; D015925:Cryopreservation; D010064:Embryo Implantation / Q000502:physiology; D005260:Female; D005615:Freezing; D006801:Humans; D019394:Ki-67 Antigen / Q000378:metabolism; D008550:Melatonin / Q000008:administration & dosage; D051379:Mice; D006080:Ovarian Follicle / Q000254:growth & development / Q000637:transplantation*; D010053:Ovary / Q000254:growth & development / Q000637:transplantation*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011247:Pregnancy; D058989:Vitrification
D019394:Ki-67 Antigen; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D008550:Melatonin
Human ovarian tissue; Ki67; Needle-immersed vitrification; PECAM; Slow-gradual freezing; TUNEL; “Improvement protocol”