Clin Exp Reprod Med.  2019 Dec;46(4):152-165. 10.5653/cerm.2019.00143.

Survival of isolated human preantral follicles after vitrification: Analyses of morphology and Fas ligand and caspase-3 mRNA expression

Affiliations
  • 1Department of Obstetrics and Gynecology, Dr. Cipto Mangunkusumo General Hospital, Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia. wiwekobudi@yahoo.co.id
  • 2Indonesian Medical Education and Research Institute (IMERI), Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia.
  • 3Department of Anatomy, IPB University, Bogor, Indonesia.
  • 4Department of Public Health, Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia.
  • 5Department of Pathology Anatomy, Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia.
  • 6Department of Biology, Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia.
  • 7Department of Histology, Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia.
  • 8Department of Obstetrics and Gynecology, Dr. Hasan Sadikin Hospital, Faculty of Medicine Universitas Padjajaran, Bandung, Indonesia.

Abstract


OBJECTIVE
This study aimed to examine the effect of vitrification on apoptosis and survival in human preantral follicles after thawing.
METHODS
This experimental study was conducted at an acute tertiary care hospital from March 2012 to April 2013. Ovaries were sliced into 5×5×1-mm pieces and divided into the following three groups: preantral follicle isolation, ovarian tissue vitrification-warming followed by follicle isolation, and immunohistochemistry of fresh ovarian tissue. For statistical analyses, the Student t-test, chi-square test, Kruskal-Wallis test, and Kaplan-Meier survival analysis were used.
RESULTS
A total of 161 preantral follicles (70% secondary) were collected from ovarian cortex tissue of six women between 30 and 37 years of age who underwent oophorectomy due to cervical cancer or breast cancer. There were no significant differences in the follicular morphology of fresh preantral follicles and vitrified follicles after thawing. The mean Fas ligand (FasL) mRNA expression level was 0.43±0.20 (relative to β-actin) in fresh preantral follicles versus 0.51±0.20 in vitrified follicles (p=0.22). The mean caspase-3 mRNA expression level in fresh preantral follicles was 0.56±0.49 vs. 0.27±0.21 in vitrified follicles (p=0.233). One vitrified-thawed secondary follicle grew and developed to an antral follicle within 6 days of culture.
CONCLUSION
Vitrification did not affect preantral follicle morphology or mRNA expression of the apoptosis markers FasL and caspase-3. Further studies are required to establish whether vitrification affects the outcomes of in vitro culture and the maturation of preantral follicles.

Keyword

Caspase-3; Fas ligand; Human preantral follicles; Vitrification

MeSH Terms

Apoptosis
Breast Neoplasms
Caspase 3*
Fas Ligand Protein*
Female
Humans*
Immunohistochemistry
In Vitro Techniques
Ovariectomy
Ovary
RNA, Messenger*
Tertiary Healthcare
Uterine Cervical Neoplasms
Vitrification*
Caspase 3
Fas Ligand Protein
RNA, Messenger
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