Submit to Animals Review for Animals Propose a Special Issue

Journal Browser

► Journal Browser

In Vitro Growth of Mammalian Follicles and Oocytes

Special Issue Editors
Special Issue Information
Keywords
Published Papers

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Animal Reproduction".

Deadline for manuscript submissions: closed (31 March 2024) | Viewed by 10966

Share This Special Issue

Special Issue Editor

Dr. Kenichiro Sakaguchi

E-Mail Website
Guest Editor

Laboratory of Veterinary Theriogenology, Joint Department of Veterinary Medicine, Faculty of Applied Biological Sciences, Gifu University, Yanagito 1-1, Gifu 501-1193, Japan
Interests: oocyte; in vitro growth (IVG); ovum-pickup–in vitro fertilization (OPU-IVF); cattle; water buffalo; ovarian reserve; heat stress

Special Issue Information

Dear Colleagues,

In mammalian ovaries, there is a large number of immature follicles, most of which are destined to degenerate before ovulation. Developing a culture system that enables small oocytes in these follicles to grow to the stage where they can be fertilized in vitro would be a significant achievement that can be of great help in assistant reproductive technology, production of farm animals, and experimental models for folliculogenesis and oogenesis.

In 2016, the generation of offspring from murine primordial germ cells using an entire in vitro growth (IVG) culture system was reported. Furthermore, the birth of pups from iPS-cell-derived oocytes differentiated in vitro became a reality in a murine model in the same year. It is expected that the application of such technology to livestock and endangered animals may no longer be just a dream in the future.

The aim of this Special Issue is to present recent research and reviews in the development of IVG of immature follicles and oocytes in mammalian species and its application to stem cell technologies and utilization in experimental models.

Dr. Kenichiro Sakaguchi
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Animals is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

in vitro growth (IVG)
pre-in vitro maturation (Pre-IVM)
in vitro differentiation (IVDi)
oocyte
follicle
ovary

Published Papers (7 papers)

Download All Papers

Editorial

Jump to: Research

4 pages, 224 KiB

Open AccessEditorial

In Vitro Growth of Mammalian Follicles and Oocytes

by Kenichiro Sakaguchi

Animals 2024, 14(9), 1355; https://doi.org/10.3390/ani14091355 - 30 Apr 2024

Viewed by 200

Abstract

Mammalian ovaries contain a large number of immature follicles, most of which are destined to degenerate before ovulation [...] Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

Research

Jump to: Editorial

14 pages, 1009 KiB

Open AccessArticle

Effect of Growth Factors and Hormones during In Vitro Growth Culture of Cumulus-Oocyte-Complexes Derived from Small Antral Follicles in Pigs

by Minji Kim, Ji-Eun Park, Yongjin Lee, Seung-Tae Lee, Geun-Shik Lee, Sang-Hwan Hyun, Eunsong Lee and Joohyeong Lee

Animals 2023, 13(7), 1206; https://doi.org/10.3390/ani13071206 - 30 Mar 2023

Cited by 1 | Viewed by 1725

Abstract

This study evaluated the effect of various growth factors and hormones in an in vitro growth (IVG) medium on the in vitro maturation (IVM) and developmental competence of oocytes derived from small antral follicles (SAFs) in pigs. Cumulus–oocyte complexes (COCs) derived from SAFs were either untreated or treated with epidermal growth factor (EGF), insulin-like factor-1 (IGF-1), insulin, or growth hormone (GH) for 2 days of IVG. Following IVG, COCs were cultured for maturation, and IVM oocytes were induced for parthenogenesis (PA). During IVG, the nuclear maturation of oocytes was significantly increased by the insulin treatment compared to other treatments. Moreover, the insulin treatment significantly increased blastocyst formation after PA relative to the No-IVG, control, EGF, and GH treatments. The cumulus expansion score after IVG-IVM was significantly higher in the insulin group than in the other groups. The glutathione (GSH) contents in IVM oocytes were increased through treatment with IGF, insulin, and GH compared to those of No-IVG oocytes. The level of reactive oxygen species (ROS) in IVM oocytes in all treatment groups was significantly lower after IVG culture than in the No-IVG group. The maturation-promoting factor (MPF) activity after IVM in the insulin-treated oocytes was significantly higher than that of the oocytes treated with EGF, IGF-1, and GH. In conclusion, this study demonstrates that insulin treatment during IVG culture improves the maturational and developmental competence of oocytes derived from SAFs in pigs through its effect on cumulus cell expansion and cytoplasmic microenvironments, such as GSH, ROS, and MPF activity. Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

► Show Figures

Figure 1

16 pages, 4319 KiB

Open AccessArticle

Relationship between Amino Acid Metabolism and Bovine In Vitro Follicle Activation and Growth

by Kenichiro Sakaguchi, Kohei Kawano, Yuki Otani, Yojiro Yanagawa, Seiji Katagiri and Evelyn E. Telfer

Animals 2023, 13(7), 1141; https://doi.org/10.3390/ani13071141 - 23 Mar 2023

Cited by 1 | Viewed by 1661

Abstract

The amino acid metabolism of bovine follicles during in vitro growth (IVG) was evaluated to identify potential indicators of health during culture. The bovine ovarian cortex was sliced, prepared as strips, and cultured for 6 days. Tissue samples were examined histologically before and after 6 days of culture, and the degree of follicle activation was classified as either high or low based on the number of growing secondary follicles present (high: 7~11; low: 0~1). In a separate experiment, secondary follicles (diameter range: 100~200 μm) were manually isolated and cultured, and their growth was monitored for 6 days. Cultured follicles were classified as growth or degenerate based on diameter change during culture (growth: +60.5~74.1 μm; degenerate: −28~15.2 μm). Free amino acids and their metabolites were measured in the spent culture medium from each group. In cultured ovarian cortical strips, the concentration of α-aminoadipic acid was significantly higher in the low activation group than in the high group (p < 0.05), while those of methionine, lysine, and arginine were higher in the high activation group. In cultured isolated secondary follicles, concentrations of methionine, tyrosine, histidine, and hydroxyproline were higher in the degenerate group (p ≤ 0.05). In conclusion, amino acid metabolism has the potential to serve as an indicator of primordial follicle activation and subsequent growth rate during bovine IVG. Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

► Show Figures

Figure 1

18 pages, 4934 KiB

Open AccessArticle

Protective Effect of Cimicifuga racemosa (L.) Nutt Extract on Oocyte and Follicle Toxicity Induced by Doxorubicin during In Vitro Culture of Mice Ovaries

by Ernando I. T. de Assis, Venância A. N. Azevedo, Miguel F. De Lima Neto, Francisco C. Costa, Laís R. F. M. Paulino, Pedro A. A. Barroso, Mariana A. M. Donato, Christina A. Peixoto, Alane P. O. Do Monte, Maria H. T. Matos, Alana N. Godinho, Jordânia M. O. Freire, Ana L. P. S. Batista, José R. V. Silva and Anderson W. B. Silva

Animals 2023, 13(1), 18; https://doi.org/10.3390/ani13010018 - 20 Dec 2022

Cited by 2 | Viewed by 1734

Abstract

This study evaluated the potential of Cimicifuga racemosa (L.) Nutt extract (CIMI) to reduce the deleterious effects of doxorubicin (DOXO) in oocytes, follicles and stromal cells in mice ovaries cultured in vitro. In experiment 1, mice ovaries were cultured in DMEM⁺ alone or supplemented with 5, 50 or 500 ng/mL CIMI, while in experiment 2, mice ovaries were cultured in DMEM⁺ alone or supplemented with 5 ng/mL CIMI (better concentration), 0.3 μg/mL DOXO or both. Thereafter, the ovaries were processed for histological (morphology, growth, activation, extracellular matrix configuration and stromal cell density), immunohistochemical (caspase-3) analyses. Follicle viability was evaluated by fluorescence microscopy (ethidium homodimer-1 and calcein) while real-time PCR was performed to analyses the levels of (mRNA for SOD, CAT and nuclear factor erythroid 2–related factor 2 (NRF2) analyses. The results showed that DOXO reduces the percentage of normal follicles and the density of stromal cells in cultured ovaries, but these harmful effects were blocked by CIMI. The DOXO reduced the percentage of primordial follicles, while the presence of CIMI alone did not influence percentage of primordial follicles. A higher staining for caspase-3 was seen in ovaries cultured in control medium alone or with DOXO when compared with those cultured with CIMI alone or both CIMI and DOXO. In addition, follicles from ovaries cultured with both CIMI and DOXO were stained by calcein, while those follicles cultured with only DOXO were stained with ethidium homodimer-1. Furthermore, ovaries cultured with CIMI or both CIMI and DOXO had higher levels of mRNA for SOD and CAT, respectively, than those cultured with only DOXO. In conclusion, the extract of CIMI protects the ovaries against deleterious effects of DOXO on follicular survival and ovarian stromal cells. Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

► Show Figures

Figure 1

12 pages, 2761 KiB

Open AccessArticle

Effects of N-acetylcysteine on Growth, Viability, and Ultrastructure of In Vitro Cultured Bovine Secondary Follicles

by Danisvânia R. Nascimento, Venância A. N. Azevedo, Pedro A. A. Barroso, Laryssa G. Barrozo, Bianca R. Silva, Anderson W. B. Silva, Mariana A. M. Donato, Christina A. Peixoto and José R. V. Silva

Animals 2022, 12(22), 3190; https://doi.org/10.3390/ani12223190 - 17 Nov 2022

Cited by 3 | Viewed by 1438

Abstract

This study aimed to investigate the effects of different concentrations of N-acetylcysteine (NAC) on the growth, antrum formation, viability, and ultrastructure of bovine secondary follicles cultured in vitro for 18 days. To this end, the follicles were cultured in TCM-199⁺ medium alone or supplemented with 1.0, 5.0, or 25.0 mM NAC. Follicular growth, antrum formation, viability (calcein-AM and ethidium homodimer-1) and ultrastructure were evaluated at the end of culture period. The results showed that 1.0 mM NAC increased the percentage of growing follicles and the fluorescence intensity for calcein-AM when compared to other treatments (p < 0.05). On the other hand, follicles cultured with 25.0 mM NAC had higher fluorescence intensity for ethidium homodimer-1, which is a sign of degeneration. Ultrastructural analysis showed that oocytes from follicles cultured in control medium alone or with 1 mM NAC had intact zonae pellucidae in close association with oolemmae, but the ooplasm showed mitochondria with a reduced number of cristae. On the other hand, oocytes from follicles cultured with 5 or 25 mM NAC had extremely vacuolated cytoplasm and no recognizable organelles. In conclusion, 1 mM NAC increases cytoplasmic calcein staining and the growth rate in bovine secondary follicles cultured in vitro, but the presence of 5 or 25 mM NAC causes damage in cellular membranes and organelles, as well as reducing the percentages of growing follicles. Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

► Show Figures

Figure 1

10 pages, 873 KiB

Open AccessArticle

Intraovarian Injection of Reconstituted Lyophilized Growth-Promoting Factor Extracted from Horse Blood Platelets (L-GF^equina) Increases Oocytes Recovery and In Vitro Embryo Production in Holstein Cows

by Silviu-Ionuț Borş, Dan-Lucian Dascălu, Alina Borş, Hossam M. Fahmy, Omaima M. Kandil and Ahmed Sabry S. Abdoon

Animals 2022, 12(19), 2618; https://doi.org/10.3390/ani12192618 - 29 Sep 2022

Cited by 2 | Viewed by 1485

Abstract

The purpose of this study was to determine the impact of intraovarian injections of a reconstituted lyophilized growth-promoting factor extracted from horse blood platelets (L-GF^equina) on the number of ovarian follicles, the recovery of cumulus–oocyte complexes (COCs), and embryo development to the blastocyst stage in Holstein cows. Thus, 12 Holstein cows were assigned to three protocols. According to the number of punctured follicles in protocol 1, ovum pick-up (OPU) was conducted on days 6 and 14 of the cycle (day 0 = estrus). In protocol 2, every large follicle (more than 7 mm) was removed, and 1 mL of L-GF^equina was intraovarian injected (day 0). Two days later, equine chorionic gonadotropin (eCG) was administered, and OPU sessions were conducted on days 6, 10, and 14. The same ovarian stimulation procedure as that in protocol 2 was performed in protocol 3, except that equine L-GF^equina was not supplied. OPU was carried out on days 6 and 10 of the cycle. The results indicate that the intraovarian injection of L-GF^equina significantly (p < 0.05) increased the number of OPU sessions per cycle, the recovery of cumulus–oocyte complexes (COCs), and the production of blastocysts. In conclusion, an intraovarian injection of L-GF^equina can improves OPU-IVEP results in Holstein cows. Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

► Show Figures

Figure 1

10 pages, 3532 KiB

Open AccessArticle

L-Carnitine Supports the In Vitro Growth of Buffalo Oocytes

by Avijit Kumar Modak, Md Hasanur Alam, Md Nuronnabi Islam, Nipa Paul, Ireen Akter, Md Abul Hashem, AKM Ahsan Kabir and Mohammad Moniruzzaman

Animals 2022, 12(15), 1957; https://doi.org/10.3390/ani12151957 - 02 Aug 2022

Viewed by 1437

Abstract

This study aimed to determine the effect of L-carnitine on the growth and subsequent nuclear maturation of buffalo small growing oocytes (92–108 µm in diameter) in vitro. Oocyte-granulosa cell complexes (OGCs) were dissected from early antral follicles of slaughtered buffaloes and cultured in in vitro growth (IVG) medium with the supplementation of different concentrations (0, 1.25, 1.875 or 2.5 mM) of L-carnitine for 6 days. The results revealed that L-carnitine increased the diameter of buffalo oocytes in vitro. The degeneration rate was significantly (p < 0.05) lower in 2.5 mM of L-carnitine-treated oocytes (10%) than others (55%, 45% and 32.5% in 0, 1.25 and 1.875 mM of L-carnitine-supplemented groups, respectively). The OGCs showed antrum-like structures significantly (p < 0.05) higher in the 2.5 mM of L-carnitine group (74.0%) than the 0- and 1.25-mM groups (34.6% and 38.1%, respectively). Furthermore, in vitro grown oocytes were placed in in vitro maturation (IVM) medium for 24 h to examine meiotic competence of in vitro grown oocytes with L-carnitine. The L-carnitine (1.875 and 2.5 mM) treated oocytes showed a higher rate of nuclear maturation up to the metaphase II (MII) stage and a lower rate of degeneration. In conclusion, L-carnitine enhances the growth, prevents degeneration, promotes the formation of antrum-like structures and supports nuclear maturation of buffalo oocytes in vitro. Full article

(This article belongs to the Special Issue In Vitro Growth of Mammalian Follicles and Oocytes)

► Show Figures