Pup birth from mouse oocytes in preantral follicles derived from vitrified and warmed ovaries followed by in vitro growth, in vitro maturation, and in vitro fertilization.
OBJECTIVE
To determine an optimum condition for vitrification of various animal species and to examine the developing capacity into pups of mouse oocytes in preantral follicles originating from cryopreserved ovaries.


METHODS
Experimental animal study.


METHODS
Laboratory environment.


METHODS
Normal (C57BL/6 x DBA2) F1 mice.


METHODS
Vitrification of animal ovaries using polyester sheets as a storage device; collection of oocyte-granulosa cell complexes by enzymatic treatment; and in vitro growth (IVG), in vitro maturation (IVM), and IVF, with embryo transplantation to pseudopregnant mice.


METHODS
Histological analysis of vitrified and warmed ovaries from several animal species; measurement of successful rates in IVG, IVM, and IVF of mouse oocytes in oocyte-granulosa cell complexes collected from the vitrified and warmed ovaries; and achievement of pup birth.


RESULTS
The vitrification method used was effective for storage of various animal ovaries. The oocytes enclosed in preantral follicles that were yielded from the vitrified-warmed ovaries preserved the capacity for developing into pups after IVG, IVM, and IVF.


CONCLUSIONS
The findings indicated that ovarian cryopreservation by vitrification and subsequent IVG, IVM, and IVF are promising methods for restoring young cancer patients' fertility. The practical information presented here is applicable to human ovarian tissues.
Hasegawa
Akiko
A
Laboratory of Developmental Biology and Reproduction, Institute for Advanced Medical Sciences, Nishinomiya, Japan.
Mochida
Nahoko
N
Ogasawara
Toshitada
T
Koyama
Koji
K
D000818:Animals; D000831:Animals, Newborn; D015925:Cryopreservation; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro; D066298:In Vitro Techniques; D008297:Male; D051379:Mice / Q000196:embryology / Q000502:physiology*; D008815:Mice, Inbred Strains; D009865:Oocytes / Q000502:physiology*; D006080:Ovarian Follicle; D010053:Ovary / Q000033:anatomy & histology; D011817:Rabbits; D013552:Swine