Optimization of the in vitro fertilization system in pigs
Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in-vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in...
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| Format: | Article |
| Language: | English |
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The Korean Society of Animal Reproduction and Biotechnology
2023-06-01
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| Series: | Journal of Animal Reproduction and Biotechnology |
| Subjects: | |
| Online Access: | https://www.e-jarb.org/journal/view.html?uid=2656&vmd=Full |
| _version_ | 1797791729684840448 |
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| author | Song-Hee Lee Xiang-Shun Cui |
| author_facet | Song-Hee Lee Xiang-Shun Cui |
| author_sort | Song-Hee Lee |
| collection | DOAJ |
| description | Background: Despite considerable technological advancements, polyspermy remains a significant challenge in in-vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs.
Methods: in-vitro-matured oocytes were co-incubated with sperm according to a modified two-step culture system.
Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 μL microdroplets (10 gametes/drop) or 500 μL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume.
Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence. |
| first_indexed | 2024-03-13T02:22:52Z |
| format | Article |
| id | doaj.art-3d0b02a0db9e4ffa9cb8818278c8bd36 |
| institution | Directory Open Access Journal |
| issn | 2671-4639 2671-4663 |
| language | English |
| last_indexed | 2024-03-13T02:22:52Z |
| publishDate | 2023-06-01 |
| publisher | The Korean Society of Animal Reproduction and Biotechnology |
| record_format | Article |
| series | Journal of Animal Reproduction and Biotechnology |
| spelling | doaj.art-3d0b02a0db9e4ffa9cb8818278c8bd362023-06-30T07:08:44ZengThe Korean Society of Animal Reproduction and BiotechnologyJournal of Animal Reproduction and Biotechnology2671-46392671-46632023-06-01382707610.12750/JARB.38.2.70Optimization of the in vitro fertilization system in pigsSong-Hee Lee0https://orcid.org/0000-0003-4233-3201Xiang-Shun Cui1https://orcid.org/0000-0001-6180-6401Department of Animal Science, Chungbuk National University, Cheongju 28644, KoreaDepartment of Animal Science, Chungbuk National University, Cheongju 28644, KoreaBackground: Despite considerable technological advancements, polyspermy remains a significant challenge in in-vitro fertilization (IVF) procedures in pigs, disrupting normal embryonic development. Here, we aimed to determine whether optimal fertilization conditions reduce the polyspermy incidence in pigs. Methods: in-vitro-matured oocytes were co-incubated with sperm according to a modified two-step culture system. Results: In the first experiment, oocytes were briefly co-incubated with sperm, washed in IVF medium, and then moved to fresh IVF medium for 5 or 6 h. Although the 6 h sperm-free cultured group had a higher penetration rate than the 5 h cultured group, the polyspermy rate significantly increased in the 6 h sperm-free cultured group. The gamete co-incubation period was either 20 or 40 min. The 40 min cultured group had a higher rate of blastocyst formation and number of total cells in blastocysts than the 20 min cultured group. In experiment 2, oocytes were inseminated with sperm separated by Pecroll treatment. Percoll treatment increased the rate of oocyte penetration and blastocyst formation compared to the control. In experiment 3, fertilized oocytes were cultured in 25 μL microdroplets (10 gametes/drop) or 500 μL (100 gametes/well) of culture medium in 4-well plates. The large volume of medium significantly reduced the number of dead oocytes and increased the rate of blastocyst formation compared to the small volume. Conclusions: Collectively, these results demonstrate that various fertilization conditions, including modified co-culture period, active sperm separation, and culture medium volume, enhance fertilization efficiency and subsequent embryonic development by decreasing polyspermy occurrence.https://www.e-jarb.org/journal/view.html?uid=2656&vmd=Fullco-culture timeembryonic developmentin-vitro fertilizationpolyspermy |
| spellingShingle | Song-Hee Lee Xiang-Shun Cui Optimization of the in vitro fertilization system in pigs Journal of Animal Reproduction and Biotechnology co-culture time embryonic development in-vitro fertilization polyspermy |
| title | Optimization of the in vitro fertilization system in pigs |
| title_full | Optimization of the in vitro fertilization system in pigs |
| title_fullStr | Optimization of the in vitro fertilization system in pigs |
| title_full_unstemmed | Optimization of the in vitro fertilization system in pigs |
| title_short | Optimization of the in vitro fertilization system in pigs |
| title_sort | optimization of the in vitro fertilization system in pigs |
| topic | co-culture time embryonic development in-vitro fertilization polyspermy |
| url | https://www.e-jarb.org/journal/view.html?uid=2656&vmd=Full |
| work_keys_str_mv | AT songheelee optimizationoftheinvitrofertilizationsysteminpigs AT xiangshuncui optimizationoftheinvitrofertilizationsysteminpigs |