Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 4 de 4
Filter
Add more filters










Database
Language
Publication year range
1.
Reprod Biol ; 17(2): 180-184, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28511995

ABSTRACT

For in vitro growth and maturation of mouse oocytes (IVG-IVM), serum is added to media up to and including the stage of oocyte maturation; this subsequently supports oocytes through fertilization and early embryo development. However, problems may occur with sera, such as batch differences and issues of biosafety. The purpose of the present study was to determine the capacity for fertilization and pre- and post-implantation development of oocytes that underwent IVG-IVM with a serum substitute. Oocyte-granulosa cell complexes from preantral follicles were cultured in medium with either fetal bovine serum (FBS), Serum Substitute Supplement™ (SSS), or Knockout™ Serum Replacement (KSR) for 10days, and were then allowed to mature for 17 h. Subsequently, more than 90% of oocytes underwent germinal vesicle breakdown (GVBD) and more than 70% reached metaphase II, with no significant difference between the groups. A lower fertilization rate, presumably due to zona hardening, was found in the serum substitute groups. Nevertheless, more than 50% of the inseminated oocytes were fertilized and 35%-45% of them underwent first cleavage and developed to the blastocyst stage. Following embryo transfer, one and four live offspring were produced from the SSS and KSR groups, respectively. The present study demonstrated that murine IVG-IVM oocytes cultured in media with a serum substitute, achieved fertilization in vitro, pre- and post-implantation development, and the delivery of live pups, although the efficiency of the process is reduced compared to FBS supplementation.


Subject(s)
In Vitro Oocyte Maturation Techniques/methods , Oocytes/physiology , Animals , Animals, Newborn , Blood Substitutes , Culture Media , Embryo Implantation/physiology , Embryo Transfer , Female , Mice , Microscopy, Electron, Transmission , Oocytes/ultrastructure , Pregnancy
2.
J Reprod Dev ; 57(6): 715-22, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21921433

ABSTRACT

This study was undertaken to examine pre- and postimplantation developmental potency of cryopreserved embryos that had undergone in vitro growth (IVG), maturation (IVM) and fertilization (IVF) of oocytes from the preantral follicle stage. An oocyte culture system for IVG and IVM was used in oocyte-granulosa cell complexes (OGCs) derived from preantral follicles in 12-day-old mice. The rate of oocyte maturation was improved by the addition of gonadotropins (FSH/LH) and cytokines (IGF-I/SCF) to culture medium for IVG. During culture for IVG, estradiol-17ß and progesterone concentrations increased progressively to the latter period of culture. This culture system enabled IVG, IVM, IVF and pre- and postimplantation development. From 90 cryopreserved 2-cell stage embryos transferred into recipients after warming, 10 live pups were produced. Cryopreservation of embryos by vitrification at the 2-cell stage showed no harmful effect on development to the blastocyst stage or on the cell numbers of the inner cell mass (ICM) and trophectoderm (TE). This study demonstrated that embryos derived from oocytes grown in vitro have tolerance for vitrification and competence to develop to term after warming. This IVG-IVM-IVF technology combined with embryo cryopreservation might be useful for assisted reproduction in mice.


Subject(s)
Cryopreservation/methods , Live Birth , Oocytes/growth & development , Ovarian Follicle/growth & development , Animals , Blastocyst/drug effects , Culture Media , Estradiol/biosynthesis , Female , Fertilization in Vitro/drug effects , Fertilization in Vitro/methods , Follicle Stimulating Hormone/pharmacology , Granulosa Cells/drug effects , Insulin-Like Growth Factor I/pharmacology , Luteinizing Hormone/pharmacology , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Oocytes/drug effects , Ovarian Follicle/drug effects , Progesterone/biosynthesis , Stem Cell Factor/pharmacology , Vitrification
3.
Hum Cell ; 22(2): 43-8, 2009 May.
Article in English | MEDLINE | ID: mdl-19379463

ABSTRACT

Under organ culture, female fetal gonads in mice cannot develop beyond the preantral follicle stage unless the follicles are individually isolated and cultured again. In this study, we investigated the effect of in vitro culture of female fetal gonads before transplantation on subsequent in vivo development. The gonads derived from female fetuses 12.5 days postcoitum were organ-cultured for 0, 7 and 14 days, and then were grafted underneath the kidney capsules of severe combined immunodeficient mice and recovered at 21, 14 and 7 days post-transplantation, respectively. The histological analysis of the grafts showed that the in vitro culture of the fetal gonads restricted follicular development to the antral follicle stage post-transplantation. In the grafts cultured for 14 days, particularly, no antral follicle was observed. However, the oocytes in these follicles had grown to around 65 microm in diameter and had competence to resume meiosis in vitro. When the fetal gonads were grafted after culture for 7 and 14 days, 13.0% and 6.8% of the oocytes progressed to the metaphase II stage, respectively. These data showed significant differences (P < 0.05) in comparison with the control group (25.3%). Our results indicate that the in vitro culture of female fetal gonads before transplantation affects the subsequent in vivo development of both follicular cells and oocytes, and in vitro oocyte maturation. However, this effect seems to be more severe in terms of follicular development when compared with oocyte growth and maturation.


Subject(s)
Oocytes/growth & development , Ovary/embryology , Animals , Female , Kidney , Male , Meiosis , Metaphase , Mice , Mice, Inbred C57BL , Mice, Inbred CBA , Organ Culture Techniques , Ovarian Follicle/growth & development , Ovary/growth & development , Ovary/transplantation , Severe Combined Immunodeficiency
4.
Hum Cell ; 17(1): 67-74, 2004 Mar.
Article in English | MEDLINE | ID: mdl-15369139

ABSTRACT

The aim of this study was to clarify the developmental and ultrastructual characteristics of oocytes grown in vitro from primordial germ cells. The female genital ridges at 12.5 days post coitus were cultured for 18 days on an insert membrane in Waymouth's MB752/1 medium, supplemented with 15% fetal bovine serum and 1 mM sodium pyruvate; subsequently, the follicles isolated from the tissue were cultured for eight days in Waymouth's medium supplemented with 5 microg/ml insulin, 5 microg/ml transferrin, 5 ng/ml selenium, 10 mIU/ml follicle stimulating hormone, and 100 ng/ml stem cell factor. The primordial germ cells developed in vitro into oocytes of more than 60 microm in diameter. The transmission electron microscopic analysis indicated that the oocytes, which developed in vitro, showed no obvious abnormality in their ultrastructure and had organelles appropriate for the oocyte size. However, a delay in the progressive changes of morphology in some of the organelles during oocyte growth was often found when comparing them to oocytes grown in vivo.


Subject(s)
Germ Cells/cytology , Oocytes/growth & development , Oocytes/ultrastructure , Animals , Cells, Cultured , Female , Meiosis , Mice , Mice, Inbred Strains , Microscopy, Electron , Oocytes/cytology , Organelles/ultrastructure
SELECTION OF CITATIONS
SEARCH DETAIL
...