Recent Advances in Reproductive Biotechnologies—Second Edition

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

Deadline for manuscript submissions: 31 October 2025 | Viewed by 2596

Special Issue Editors


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Guest Editor
Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Department of Agricultural Sciences and Natural Resources, Faculty of Agriculture and Forestry Sciences, Universidad de La Frontera, Temuco, Chile
Interests: in vitro embryo production; early embryo development; embryonic pluripotency; developmental epigenetics; micromanipulation of gametes and embryos; ICSI; SCNT
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Guest Editor
Instituto Nacional de Investigação Agrária e Veterinária, Quinta da Fonte Boa, 2005-048 Vale de Santarém, Portugal
Interests: small ruminants; reproduction physiology and biotechnologies; andrology; reproductive efficiency and methods; reproduction parameters; sperm cryopreservation (new extenders, tecniques, new methods, etc.); cervical artificial insemination
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Recently, there have been increasing advances in “Reproductive Biotechnologies” which seek to optimize reproductive efficiency, productivity,  gamete selection, and preservation and health performance under current challenging environmental conditions. During recent decades, reproductive technologies have opened new avenues for studying and manipulating reproductive biology in different mammalian species with important economical, societal, and medical implications. In human medicine, one of the main applications is the treatment of infertility, but in animal reproduction, there are other applications. Among these are wildlife and domestic cell conservation, the improvement and preservation of animal genetics, and the enhancement of reproductive efficiency. Reproductive biotechnologies include artificial insemination, embryo transfer, estrus synchronization and superovulation, multiple ovulation embryo transfer, laparoscopic ovum pick-up, laparoscopic insemination, the in vitro production of embryos, intracytoplasmic sperm injection, the refrigeration and cryopreservation of sperm, the cryopreservation of oocytes and embryos, the sexing of sperm and embryos, embryo splitting, cloning and gene transfer, the production of artificial gametes and embryos, and marker-assisted selection. This is why we are pleased to invite you to publish high-quality research papers, review articles, and communications to this Special Issue, addressing new advances in the field of reproductive biotechnologies in mammals.

The research topics may include (but are not limited to) the following: reproductive health; infertility; early embryo development and pregnancy; progress in molecular markers associated with fertility, gamete and embryo quality, and reproductive disorders, as well as designing breeding strategies; assisted reproductive technologies such as cryopreservation, in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), somatic cell nuclear transfer (SCNT), the micromanipulation of gametes and embryos, the genetic and genomic analysis of gametes and embryos, embryo biopsy, embryo transfer, fertility preservation, artificial insemination, oocyte and embryo culture, genomic selection and manipulation, transgenesis, transcriptomics and epigenomics, embryo outgrowths, and embryonic stem cells; and the improvement of methods and techniques for gamete evaluation and reproductive efficiency methods.

Original, high-quality contributions that are not yet published or under review by other journals are sought. We hope that this Issue will provide novel insights into the recent advancements in the field of reproductive biotechnologies, as well in basic and applied reproduction science.

We look forward to receiving your contributions.

Dr. Luis Águila Paredes
Dr. João Pedro Barbas
Guest Editors

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

  • reproductive biotechnologies
  • productivity
  • embryo development
  • andrology
  • sperm cryopreservation

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Published Papers (3 papers)

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Research

13 pages, 2029 KiB  
Article
Human Recombinant Interleukin-6 and Leukemia Inhibitory Factor Improve Inner Cell Mass Cell Number but Lack Cryoprotective Activities on In Vitro-Produced Bovine Blastocysts
by Mary A. Oliver, Kayla J. Alward, Michelle L. Rhoads and Alan D. Ealy
Animals 2025, 15(5), 668; https://doi.org/10.3390/ani15050668 - 25 Feb 2025
Viewed by 513
Abstract
This work explored whether supplementing recombinant human interleukin-6 (IL6), interleukin-11 (IL11), or leukemia inhibitory factor (LIF) improves IVP bovine embryo development, morphology, and cryosurvivability. Embryos were treated from day 5 to 8 post-fertilization with either the carrier only (control) or 100 ng/mL of [...] Read more.
This work explored whether supplementing recombinant human interleukin-6 (IL6), interleukin-11 (IL11), or leukemia inhibitory factor (LIF) improves IVP bovine embryo development, morphology, and cryosurvivability. Embryos were treated from day 5 to 8 post-fertilization with either the carrier only (control) or 100 ng/mL of IL6, IL11, or LIF. Blastocyst formation and stage were determined on day 7 and 8. A subset of day 8 blastocysts was processed for immunofluorescence to count trophectoderm (TE) and inner cell mass (ICM) cell numbers and another subset was slow frozen and stored in liquid nitrogen until thawing. No differences in the blastocyst rate or blastocyst stage of development were detected. Increases in ICM cell numbers were observed for IL6 and LIF but not the IL11 treatment. None of the cytokine treatments applied before freezing affected post-thaw survival, TE or ICM cell number, or cell death 24 h after thawing. In conclusion, supplementing IL6 and LIF improves ICM cell numbers in non-frozen blastocysts, but there was no evidence that any of these cytokine treatments contain cryoprotective properties in bovine embryos. Full article
(This article belongs to the Special Issue Recent Advances in Reproductive Biotechnologies—Second Edition)
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14 pages, 3501 KiB  
Article
Achieving Optimal Transfection Conditions in Chicken Primordial Germ Cells Under Feeder- and Serum-Free Medium
by Zhifeng Zhao, Xian Zou, Ying Zhu, Yanhua He, Endashaw Jebessa, Jiannan Zhang, Jian Ji, Peng Chen and Chenglong Luo
Animals 2025, 15(4), 590; https://doi.org/10.3390/ani15040590 - 18 Feb 2025
Viewed by 523
Abstract
The successful application of primordial germ cells (PGCs) is an ideal method for generating gene-edited birds. However, barriers to efficient DNA transfection in PGCs lead to low transfection efficiency, limiting the generation of genetically modified chickens. The current study utilized chemical transfection and [...] Read more.
The successful application of primordial germ cells (PGCs) is an ideal method for generating gene-edited birds. However, barriers to efficient DNA transfection in PGCs lead to low transfection efficiency, limiting the generation of genetically modified chickens. The current study utilized chemical transfection and electroporation methods to determine the optimal transfection conditions for the PGC line under feeder- and serum-free medium. Among the tested methods, the Lonza electroporation system exhibited the highest transduction efficiency, with a previously unreported rate of 71.13 ± 1.26%. Optimal transfection conditions were achieved using 4 µg of DNA and 100 µL of EntransterTM-E in 1 × 106 PGCs. Furthermore, the optimal electroporation conditions resulted in low cell death and normal expression of pluripotency-related genes, highlighting the low cytotoxicity. The resulting electroporation models were then used to deliver the enhanced green fluorescent protein (EGFP) gene to the Z chromosome with a Cas9-gRNA plasmid, achieving a 7-day insertion efficiency of 14.63 ± 1.07%. Our study highlights the vast potential of electroporation technology for the transfection of PGCs. Full article
(This article belongs to the Special Issue Recent Advances in Reproductive Biotechnologies—Second Edition)
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13 pages, 3047 KiB  
Article
Comparison of Development and Antioxidative Ability in Fertilized Crossbred (Yorkshire × Landrace × Duroc) Oocytes Using Duroc and Landrace Sperm
by Hayoung Lee, Hyewon Kim, Jisoon An, Hee-Tae Cheong and Sang-Hee Lee
Animals 2024, 14(24), 3562; https://doi.org/10.3390/ani14243562 - 10 Dec 2024
Viewed by 947
Abstract
Pig production through crossbreeding methods is a pillar of the swine industry; however, research on the fertilization ability of male pigs in crossbreeds is lacking. Therefore, this study investigated the effects of Duroc sperm (DS) and Landrace sperm (LS) on fertility in Yorkshire [...] Read more.
Pig production through crossbreeding methods is a pillar of the swine industry; however, research on the fertilization ability of male pigs in crossbreeds is lacking. Therefore, this study investigated the effects of Duroc sperm (DS) and Landrace sperm (LS) on fertility in Yorkshire × Landrace × Duroc (YLD) oocytes. Sperm were collected from the Duroc and Landrace species, and sperm characteristics, viability, and acrosome reactions were analyzed using flow cytometry. Oocytes were collected from YLD ovaries, and the fertility of DS and LS was determined using in vitro fertilization (IVF). Reactive oxygen species (ROS) and antioxidative abilities were analyzed using H2DCFDA and a Cell Tracker Red assay. Pluripotency (OCT4, SOX2, and NANOG), antioxidative (SOD1, SOD2, CAT, and GPx1), apoptotic (Bax and Bcl-2), and cell cycle-related (Cdc2 and CCNB1) genes were detected using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in oocytes fertilized with sperm. The results showed no significant difference in viability or acrosome reaction between DS and LS. ROS levels were significantly lower in the LS group than in the DS group, whereas glutathione (GSH) levels in the embryo did not significantly differ between the DS and LS groups. The OCT4, GPx1, and Cdc2 mRNA expression levels were significantly higher in the LS than DS groups. Blastocyst formation was significantly higher in the LS than DS groups. ROS levels were reduced, and blastocyte formation was increased in LS-obtained embryos. In conclusion, these results provide a fundamental understanding of using Landrace semen in the three-way crossbreeding of YLD pigs. Full article
(This article belongs to the Special Issue Recent Advances in Reproductive Biotechnologies—Second Edition)
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