The effect of leukemia inhibitory factor and coculture on the in vitro maturation and ultrastructure of vitrified and nonvitrified isolated mouse preantral follicles.
OBJECTIVE
To develop a system for in vitro maturation of preantral follicles isolated from vitrified and nonvitrified mouse ovaries by leukemia inhibitory factor (LIF) and coculture with cumulus cells.


METHODS
Experimental study.


METHODS
University-based research laboratory.


METHODS
Twelve- to 14-day-old National Medical Research Institute female mice.


METHODS
Vitrification of mouse ovaries and in vitro maturation of follicles.


METHODS
The growth, maturation, steroidogenesis, alkaline phosphatase activity, and ultrastructure of preantral follicles derived from vitrified and nonvitrified mouse ovaries and the developmental capacity of embryos obtained from metaphase II to blastocyst stage.


RESULTS
The follicular diameters were increased in the presence of LIF in the simple culture, coculture, and vitrified coculture groups. The survival and developmental rates of follicles were not significantly different between groups. The E(2) production was increased in all groups during the culture period. Alkaline phosphatase activity was observed in the follicles of cryosectioned slices and cultured preantral follicles. There was no remarkable change in ultrastructural maturation features of cultured follicles.


CONCLUSIONS
LIF alone or in combination with the coculture system increased the growth of cultured preantral follicles but had no effect on their maturation; the ultrastructure, growth, and maturation of vitrified follicles were the same in the cultured groups and in the controls.
Haidari
Kamran
K
Anatomy Department, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Salehnia
Mojdeh
M
Rezazadeh Valojerdi
Mojtaba
M
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002450:Cell Communication / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D015925:Cryopreservation; D054885:Cumulus Cells / Q000378:metabolism*; D046149:Embryo Culture Techniques; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D051379:Mice; D006080:Ovarian Follicle / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D013084:Sperm-Ovum Interactions / Q000187:drug effects; D014021:Tissue Preservation / Q000379:methods*
D053605:Leukemia Inhibitory Factor; D004958:Estradiol; D000469:Alkaline Phosphatase