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Comparative Study
. 2021 Oct 29;67(5):327-331.
doi: 10.1262/jrd.2021-091. Epub 2021 Sep 5.

Optimal conditions for mouse follicle culture

Saya Ota  1 , Shinya Ikeda  1 , Tomoya Takashima  1 , Yayoi Obata  1
Affiliations
Comparative Study

Optimal conditions for mouse follicle culture

Saya Ota et al. J Reprod Dev. .

Abstract

Mammalian ovaries contain a large number of immature follicles. Follicular culture can contribute to the production of fertile oocytes from latent immature follicles, providing a useful tool for exploring the developmental competencies and related factors that oocytes acquire during growth. However, the potential of oocytes produced by follicular culture is limited. Herein, the optimal follicular culture conditions for the addition of polyvinylpyrrolidone to the medium and oxygen concentration were investigated. Polyvinylpyrrolidone with a high molecular weight (≥ 360,000) and a 7% oxygen concentration were found to increase the blastocyst formation rate by more than 20% compared with conventional culture conditions. Although the developmental ability of oocytes produced by follicular culture remained inferior to that of in vivo-derived oocytes, these findings may pave the way for enhanced production of fertile oocytes in vitro and for studying the process of full developmental potency acquisition by oocytes.

Keywords: Culture; Developmental competence; Oocyte; Oocyte growth.

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Conflict of interest statement

The authors declare that there are no conflicts of interests.

Figures

Fig. 1.
Fig. 1.
Meiotic and developmental competence of oocytes produced by follicular culture (FC) with medium containing polyvinylpyrrolidone (PVP) compounds of various molecular weights (MW). (A) Percentage of oocytes that reached the second meiosis stage after FC and in vitro maturation (IVM). Error bars indicate the standard deviation. Statistical significance was analyzed by Tukey’s test. Different letters represent a significant difference (P < 0.01). (B) Percentage of normally fertilized oocytes that developed into each stage after FC, IVM, and in vitro fertilization. Statistical significance was analyzed by Tukey’s test. Different letters represent a significant difference (P < 0.01). (C) Viscosity and osmolality of the medium containing various PVP compounds.
Fig. 2.
Fig. 2.
Meiotic and developmental competence of oocytes produced by follicular culture (FC) under various oxygen concentrations. (A) Conditions of FC, in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC) of embryos tested in this study. (B) Percentage of oocytes that reached the second meiosis stage after FC and IVM. Error bars indicate the standard deviation. Statistical significance was analyzed by Tukey’s test. Different letters represent a significant difference (a vs. bc, a vs. d, ab vs. d, P < 0.01; bc vs. d, P < 0.05). (C) Percentage of normally fertilized oocytes that developed into each stage after FC, IVM, and IVF. Error bars indicate the standard deviation. Statistical significance was analyzed by Tukey’s test. Different letters represent a significant difference (P < 0.01). (D) Numbers of OCT4+ and CDX2+ cells in the resultant blastocysts. (E, F) Immunostaining images of blastocysts produced under (E) 7% and (F) 20% O2 conditions using anti-OCT4 (green) and anti-CDX2 (magenta) antibodies. Nuclei (blue) were counterstained with DAPI. Bars indicate 20 µm.
See this image and copyright information in PMC

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