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BOR - Papers in Press, published online ahead of print December 11, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.013029
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BIOLOGY OF REPRODUCTION 68, 1682–1686 (2003)
DOI: 10.1095/biolreprod.102.013029
© 2003 by the Society for the Study of Reproduction, Inc.

Gamete Biology

A Revised Protocol for In Vitro Development of Mouse Oocytes from Primordial Follicles Dramatically Improves Their Developmental Competence1

Marilyn J. O'Brien, Janice K. Pendola, and John J. Eppig2

The Jackson Laboratory, Bar Harbor, Maine 04609

The objective of this study was to improve the conditions for oocyte development in vitro beginning with the primordial follicles of newborn mice. Previous studies showed that oocytes competent of meiotic maturation, fertilization, and preimplantation could develop in vitro from primordial follicles. However, the success rates were low and only one live offspring was produced (0.5% of embryos transferred). A revised protocol was compared with the original protocol using oocyte maturation and preimplantation development as end points. The percentage of oocytes maturing to metaphase II and developing to the blastocyst stage was significantly improved using the revised protocol. In addition, we compared the production of offspring from two-cell stage embryos derived from in vitro-grown and in vivo-grown oocytes. Of 1160 transferred two-cell stage embryos derived from in vitro-grown oocytes, 66 (5.7%) developed to term and 7 pups (10.6%) died at birth. The remaining 59 pups (27 females, 32 males) survived to adulthood. By comparison, of 437 transferred two-cell stage embryos derived from in vivo-grown oocytes, 76 (17.4%) developed to term and 4 (5.3%) died at birth. The remaining 72 pups (35 females, 37 males) survived to adulthood. These studies provide proof of the principle that fully competent mammalian oocytes can develop in vitro from primordial follicles and present a significant advance in oocyte culture technology.

1 This research was supported by grant HD21970 (J.J.E.) from the National Institute for Child Health and Human Development. J.K.P. was supported by National Institutes of Health grant NRSA F32 HD42373. Scientific Resources at The Jackson Laboratory are supported in part by a Cancer Center Core Grant (CA34194) from the National Cancer Institute.

2 Correspondence. FAX: 207 288 6073; jje{at}jax.org






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Copyright © 2003 by the Society for the Study of Reproduction.