Effect of Droplet Size and Number of Oocytes Examined on Mouse Oocyte Quality in In Vitro Maturation

Nishio, Manami; Hoshino, Yumi; Sato, Eimei

doi:10.1274/jmor.28.53

Cited by 6 publications

(11 citation statements)

References 19 publications

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“…In fact, Nishio et al. () reported that the nuclear maturation rate using monoculture with CD is significantly lower than when using coculture with CD. Usually, in oocyte culture systems, more than one oocyte per culture medium drop has been used in IVM.…”

Section: Discussionmentioning

confidence: 99%

“…It has previously been thought that the nuclear maturation rate is low in IVM by monoculture of oocytes as compared with coculture. In fact, Nishio et al (2011) reported that the nuclear maturation rate using monoculture with CD is significantly lower than when using coculture with CD. Usually, in oocyte culture systems, more than one oocyte per culture medium drop has been used in IVM.…”

Section: Discussionmentioning

confidence: 99%

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Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes

Ishikawa

Machida

Hiraga

et al. 2014

Reprod Domestic Animals

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We analysed the effect of three antioxidants that have different functional mechanisms on the in vitro maturation (IVM) of porcine oocytes. Single oocyte monoculture using the hanging drop (HD) system has some advantages such as improving analysis efficiency brought by the smaller number of samples than the number of oocytes cultured in one drop. Direct effects of ligands on single oocytes could also be detected without considering the effects of paracrine factors from other oocytes. After 22 h of pre-culture, denuded oocytes were cultured for 22 h with 0.01 and 0.1 μg/ml of L-carnitine (LC), lactoferrin (LF) or sulforaphane (SF) in the presence/non-presence of oxidant stress induced by H2O2 supplementation to evaluate the reducing effects against oxidative stress on nuclear maturation. As a result, compared with LC and SF, LF showed effective reduction in oxidative stress at a lower concentration (0.01 μg/ml), suggesting that LF is a more effective antioxidant in porcine oocyte IVM. Additionally, LF also increased maturation rate even in culture without H2O2. Our results clearly suggest that the HD monoculture system is useful for screening the substances that affect porcine oocyte culture.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes

Ishikawa

Machida

Hiraga

et al. 2014

Reprod Domestic Animals

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“…Previously, we have investigated the influence of droplet size and number of COCs in the IVM experiment on mouse oocyte quality [6]. Our results demonstrated no significant difference between the groups with 20 COCs/100 lL and five COCs/200 lL, and we suggest that B5 COCs in the IVM culture produces greater blastocyst numbers.…”

Section: Introductionmentioning

confidence: 48%

“…In our previous study, we used the IVM culture drop (CD) method [6]. In this method, a microdroplet is placed on the bottom of a dish under oil.…”

Section: Introductionmentioning

confidence: 99%

Effect of single‐oocyte culture system on in vitro maturation and developmental competence in mice

Nishio

Hoshino

Tanemura

et al. 2014

Reprod Medicine & Biology

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Purpose To investigate whether single-culture systems influence the quality of in vitro-matured oocytes, we examined the maturation and developmental competence of oocytes obtained by grouped in vitro maturation (IVM) or single IVM. Methods In vitro-matured oocytes were obtained using the culture drop (CD) method for the grouped IVM experiments, and the CD and hanging drop (HD) method for the single IVM experiments. To evaluate oocyte developmental competence, we performed in vitro fertilization and culture, and counted the number of blastocysts. To evaluate the oocyte cytoplasmic maturation, we measured the maturation promoting factor (MPF) expression levels. Results Oocytes cultured singly had lower maturity and developmental competence than the grouped IVM oocytes. However, enhanced oocyte fertility and blastocyst quality was achieved by the HD single IVM method. Additionally, the MPF activity level increased in all culture methods, compared to the control; however, it lagged behind nuclear maturation. Conclusions These results suggest that the HD method is efficient for single IVM.

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“…The components media in the different volume media (50, 100 and 200 mL/droplet medium) would sufficient amount for oocytes maturation. Oocytes maturation rates were higher when oocytes were cultured in the 50, 100 and 200 mL/droplet medium than in the 20 mL/droplet medium [26].…”

Section: Discussionmentioning

confidence: 84%

BMP-15 m-RNA expression of mouse oocytes in vitro maturation in different droplet medium volume

Rahayu

Widodo

Hoshino

et al. 2015

Asian Pacific Journal of Reproduction

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Keywords: BMP-15 m-RNA Droplet medium volume In vitro maturation Mouse Oocytes A B S T R A C T Objective: To investigate droplet medium volume effect on the BMP-15 m-RNA expression. Methods: Oocytes are collected from mice ovaries by puncturing with a sterile 26-G needle. The droplet medium volumes are using 50 mL, 100 mL and 200 mL. The BMP-15 m-RNA expression is determined in each group. Results: The results indicated that BMP-15 m-RNA expression did not significantly difference when oocyte were cultured in 50 and 100 mL/droplet medium volume, but significantly difference (P < 0.05) when oocytes were cultured in 200 mL/droplet medium volume. Conclusions: The highest BMP-15 m-RNA expression occurs when oocytes are cultured in 200 mL/droplet medium volume.

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Effect of Droplet Size and Number of Oocytes Examined on Mouse Oocyte Quality in In Vitro Maturation

Cited by 6 publications

References 19 publications

Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes

Hanging Drop Monoculture for Selection of Optimal Antioxidants During In Vitro Maturation of Porcine Oocytes

Effect of single‐oocyte culture system on in vitro maturation and developmental competence in mice

BMP-15 m-RNA expression of mouse oocytes in vitro maturation in different droplet medium volume

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