Conditions to optimise the developmental competence of immature equine oocytes*
Elizabeth S. Metcalf
A B G , Keith R. Masterson B , David Battaglia B , Jeremy G. Thompson C , Robert Foss D , Richard Beck E , Nancy L. Cook F and Thomas O’Leary B
+ Author Affiliations
- Author Affiliations
A Honahlee PC, 14005 SW Tooze Road, Sherwood, OR 97140, USA.
B Andrology Division, Department of Obstetrics and Gynecology, Oregon Health and Science University School of Medicine, 3303 S. Bond Avenue, Portland, OR 97239, USA.
C School of Medicine, The University of Adelaide, Medical School, South Adelaide, SA 5005, Australia.
D Equine Medical Services, 5851 E Deer Park Road, Columbia, MO 65201, USA.
E In Foal, Inc., 39185 Diamond Valley Road, Hemet, CA 92543, USA.
F Advanced Equine Reproduction, 1145 Arroyo Mesa, Solvang, CA 93463, USA.
G Corresponding author. Email: honahlee@imagina.com
Reproduction, Fertility and Development 32(11) 1012-1021 https://doi.org/10.1071/RD19249
Submitted: 15 July 2019 Accepted: 11 June 2020 Published: 9 July 2020
Abstract
Optimising the developmental potential of immature equine oocytes and in vitro-produced (IVP) embryos was explored through modifications of established media and holding temperature. In Experiment 1, delaying spontaneous resumption of meiosis through the process of simulated physiological oocyte maturation with the addition of the adenylate cyclase activator forskolin (50 µM) and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (100 µM) to overnight holding medium before maturation improved blastocyst production (P < 0.05). In Experiment 2, the blastocyst production rate was increased significantly when cumulin (100 ng mL−1) was added to the overnight holding or culture media (P < 0.05). In Experiment 3, immature oocytes held overnight at 16°C before maturation had improved developmental competence than those held at 20°C and 5°C (P < 0.05). There was no difference between maturation rates, but blastocyst formation per cleaved oocyte was significantly greater in oocytes held overnight at 16°C than at 20°C or 5°C. Furthermore, blastocyst formation per recovered oocyte and per fertilised oocyte was greater when oocytes were held before maturation at 16°C than at 5°C (P < 0.05). In Experiment 4, the addition of sodium ascorbate (AC; 50 µg mL−1) to the maturation and/or culture media of oocytes and IVP embryos did not improve blastocyst production, but did appear to lower cleavage rates compared with oocytes and embryos cultured without AC.
Additional keywords: blastocyst, cell culture, intracytoplasmic sperm injection, IVF, IVM.
References
Ahmadi, E., Shirazi, A., Shams-Esfandabadi, N., and Nazari, H. (2019). Antioxidants and glycine can improve the developmental competence of vitrified/warmed ovine immature oocytes. Reprod. Domest. Anim. 54, 595–603.| Antioxidants and glycine can improve the developmental competence of vitrified/warmed ovine immature oocytes.Crossref | GoogleScholarGoogle Scholar | 30637807PubMed |
Albuz, F. K., Sasseville, M., Lane, M., Armstrong, D. T., Thompson, J. G., and Gilchrist, R. B. (2010). Simulated physiological oocyte maturation (SPOM): a novel in vitro maturation system that substantially improves embryo yield and pregnancy outcomes. Hum. Reprod. 25, 2999–3011.
| Simulated physiological oocyte maturation (SPOM): a novel in vitro maturation system that substantially improves embryo yield and pregnancy outcomes.Crossref | GoogleScholarGoogle Scholar | 20870682PubMed |
Ambruosi, B., Lacalandra, G. M., Iorga, A. I., De Santis, T., Mugnier, S., and Matarrese, R. (2009). Cytoplasmic lipid droplets and mitochondrial distribution in equine oocytes: implications on oocyte maturation, fertilization and developmental competence after ICSI. Theriogenology 71, 1093–1104.
| Cytoplasmic lipid droplets and mitochondrial distribution in equine oocytes: implications on oocyte maturation, fertilization and developmental competence after ICSI.Crossref | GoogleScholarGoogle Scholar | 19167745PubMed |
Behrouzi, A., Colazo, M. G., and Ambrose, D. J. (2016). Alterations in bone morphogenetic protein 15, growth differentiation factor 9, and gene expression in granulosa cells in preovulatory follicles of dairy cows given porcine LH. Theriogenology 85, 1249–1257.
| Alterations in bone morphogenetic protein 15, growth differentiation factor 9, and gene expression in granulosa cells in preovulatory follicles of dairy cows given porcine LH.Crossref | GoogleScholarGoogle Scholar | 26794084PubMed |
Buell, M., Chitwood, J. L., and Ross, P. (2015). cAMP modulation during sheep in vitro oocyte maturation delays progression of meiosis without affecting oocyte parthenogenetic developmental competence. Anim. Reprod. Sci. 154, 16–24.
| cAMP modulation during sheep in vitro oocyte maturation delays progression of meiosis without affecting oocyte parthenogenetic developmental competence.Crossref | GoogleScholarGoogle Scholar | 25595334PubMed |
Carnevale, E. M., Maclellan, L. J., Coutinho da Silva, M. A., and Squires, E. L. (2003). Pregnancies attained after collection and transfer of oocytes from ovaries of five euthanatized mares. J. Am. Vet. Med. Assoc. 222, 60–62.
| Pregnancies attained after collection and transfer of oocytes from ovaries of five euthanatized mares.Crossref | GoogleScholarGoogle Scholar | 12523482PubMed |
Choi, Y. H., Love, L. B., Varner, D. D., and Hinrichs, K. (2006). Holding immature oocytes in the absence of meiotic inhibitors: effect of germinal vesicle chromatin and blastocyst development after intracytoplasmic sperm injection. Theriogenology 66, 955–963.
| Holding immature oocytes in the absence of meiotic inhibitors: effect of germinal vesicle chromatin and blastocyst development after intracytoplasmic sperm injection.Crossref | GoogleScholarGoogle Scholar | 16574209PubMed |
Choi, Y. H., Ritthaler, J., and Hinrichs, K. (2014). Production of a mitochondrial-DNA identical cloned foal using oocytes recovered from immature follicles of selected mares. Theriogenology 82, 411–417.
| Production of a mitochondrial-DNA identical cloned foal using oocytes recovered from immature follicles of selected mares.Crossref | GoogleScholarGoogle Scholar | 24888683PubMed |
Dalvit, G., Llanes, S. P., Descalzo, A., Insani, M., Beconi, M., and Cetica, P. (2005). Effect of alpha-tocopherol and ascorbic acid on bovine oocyte in vitro maturation. Reprod. Domest. Anim. 40, 93–97.
| Effect of alpha-tocopherol and ascorbic acid on bovine oocyte in vitro maturation.Crossref | GoogleScholarGoogle Scholar | 15819954PubMed |
Diaw, M., Salgado, R. M., Canesin, H. S., Gridley, N., and Hinrichs, K. (2018). Effect of different shipping temperatures (~22°C vs. ~7°C) and holding media on blastocyst development after overnight holding of immature equine cumulus–oocyte complexes. Theriogenology 111, 62–68.
| Effect of different shipping temperatures (~22°C vs. ~7°C) and holding media on blastocyst development after overnight holding of immature equine cumulus–oocyte complexes.Crossref | GoogleScholarGoogle Scholar | 29428846PubMed |
Dini, P., Bogado Pascottini, O., Ducheyne, K., Hostens, M., and Daels, P. (2016). Holding equine oocytes in a commercial embryo-holding medium: new perspective on holding temperature and maturation time. Theriogenology 86, 1361–1368.
| Holding equine oocytes in a commercial embryo-holding medium: new perspective on holding temperature and maturation time.Crossref | GoogleScholarGoogle Scholar | 27268297PubMed |
Ducibella, T., Anderson, E., Albertini, D. F., Aalberg, J., and Rangarajan, S. (1988). Quantitative studies of changes in cortical granule number and distribution in the mouse oocyte during meiotic maturation. Dev. Biol. 130, 184–197.
| Quantitative studies of changes in cortical granule number and distribution in the mouse oocyte during meiotic maturation.Crossref | GoogleScholarGoogle Scholar | 3141231PubMed |
Ezoe, K., Yabuuchi, A., Tani, T., Mori, C., Miki, T., Takayama, Y., Beyhan, Z., Kato, Y., Okuno, T., Kobayashi, T., and Kato, K. (2015). Developmental competence of vitrified–warmed bovine oocytes at the germinal-vesicle stage is improved by cyclic adenosine monophosphate modulators during in vitro maturation. PLoS One 10, e0126801.
| Developmental competence of vitrified–warmed bovine oocytes at the germinal-vesicle stage is improved by cyclic adenosine monophosphate modulators during in vitro maturation.Crossref | GoogleScholarGoogle Scholar | 25965267PubMed |
Farghaly, T., Khalifa, E., Mostafa, S., Hussein, M., Bedaiwy, M., and Ahmady, A. (2015). The effect of temporary meiotic attenuation on the in vitro maturation outcome of bovine oocytes. In vitro Cell. Dev. Biol. Anim. 51, 662–671.
| The effect of temporary meiotic attenuation on the in vitro maturation outcome of bovine oocytes.Crossref | GoogleScholarGoogle Scholar | 25784604PubMed |
Ferré-Pujol, P., Nguyen, X. K., Nagahara, T., Bui, T. T. M., Wakai, T., and Funahashi, H. (2019). Removal of cumulus cells around 20 h after the start of in vitro maturation improves the meiotic competence of porcine oocytes via reduction in cAMP and cGMP levels. J. Reprod. Dev. 65, 177–182.
| Removal of cumulus cells around 20 h after the start of in vitro maturation improves the meiotic competence of porcine oocytes via reduction in cAMP and cGMP levels.Crossref | GoogleScholarGoogle Scholar | 30745497PubMed |
Ferreira, E. M., Vireque, A. A., Adona, P. R., Meirelles, F. V., Ferriani, R. A., and Navarro, P. A. (2009). Cytoplasmic maturation of bovine oocytes: structural and biochemical modifications and acquisition of developmental competence. Theriogenology 71, 836–848.
| Cytoplasmic maturation of bovine oocytes: structural and biochemical modifications and acquisition of developmental competence.Crossref | GoogleScholarGoogle Scholar | 19121865PubMed |
Foss, R., Ortis, H., and Hinrichs, K. (2013). Effect of potential oocyte transport protocols on blastocyst rates after intracytoplasmic sperm injection in the horse. Equine Vet. J. Suppl. 45, 39–43.
| Effect of potential oocyte transport protocols on blastocyst rates after intracytoplasmic sperm injection in the horse.Crossref | GoogleScholarGoogle Scholar |
Gérard, N., and Robin, E. (2019). Cellular and molecular mechanisms of the preovulatory follicle differentiation and ovulation: what do we know in the mare relative to other species. Theriogenology 130, 163–176.
| Cellular and molecular mechanisms of the preovulatory follicle differentiation and ovulation: what do we know in the mare relative to other species.Crossref | GoogleScholarGoogle Scholar | 30921545PubMed |
Gilchrist, R. B., Zeng, X., Wang, H. T., Richani, D., Smitz, J., and Thompson, J. G. (2015). Reevaluation and evolution of the simulated physiological oocyte maturation system. Theriogenology 84, 656–657.
| Reevaluation and evolution of the simulated physiological oocyte maturation system.Crossref | GoogleScholarGoogle Scholar | 25958085PubMed |
Gilchrist, R. B., Luciano, A. M., Richani, D., Zeng, H. T., Wang, X., Vos, M. D., Sugimura, S., Smitz, J., Richard, F. J., and Thompson, J. G. (2016). Oocyte maturation and quality: role of cyclic nucleotides. Reproduction 152, R143–R157.
| Oocyte maturation and quality: role of cyclic nucleotides.Crossref | GoogleScholarGoogle Scholar | 27422885PubMed |
Gomes, R. G., Lisboa, L. A., Silva, C. B., Max, M. C., Marino, P. C., Oliveira, R. L., González, S. M., Barreiros, T. R., Marinho, L. S., and Seneda, M. M. (2015). Improvement of development of equine preantral follicles after 6 days of in vitro culture with ascorbic acid supplementation. Theriogenology 84, 750–755.
| Improvement of development of equine preantral follicles after 6 days of in vitro culture with ascorbic acid supplementation.Crossref | GoogleScholarGoogle Scholar | 26074067PubMed |
Goudet, G., Bezard, J., Duchamp, G., Gerard, N., and Palmer, E. (1997). Equine oocyte competence for nuclear and cytoplasmic in vitro maturation: effect of follicle size and hormonal environment. Biol. Reprod. 57, 232–245.
| Equine oocyte competence for nuclear and cytoplasmic in vitro maturation: effect of follicle size and hormonal environment.Crossref | GoogleScholarGoogle Scholar | 9241036PubMed |
Guimarães, A. L., Pereira, S. A., Leme, L. O., and Dode, M. A. (2015). Evaluation of the simulated physiological oocyte maturation system for improving bovine in vitro embryo production. Theriogenology 83, 52–57.
| Evaluation of the simulated physiological oocyte maturation system for improving bovine in vitro embryo production.Crossref | GoogleScholarGoogle Scholar | 25447152PubMed |
Guimarães, A. L., Pereira, S. A., Diógenes, M. N., and Dode, M. A. (2016). Effect of insulin–transferrin–selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation on in vitro bovine embryo development. Zygote 24, 890–899.
| Effect of insulin–transferrin–selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation on in vitro bovine embryo development.Crossref | GoogleScholarGoogle Scholar | 27748220PubMed |
Hawley, L. R., Enders, A. C., and Hinrichs, K. (1995). Comparison of equine and bovine oocyte–cumulus morphology within the ovarian follicle. In ‘Equine Reproduction VI.’ (Eds. D. C. Sharp and F. W. Bazer.) pp. 243–252. (Society for the Study of Reproduction: Madison.)
Leal, G. R., Monteiro, C. A. S., Souza-Fabjan, J. M. C., Vasconcellos, C. O. P., Nogueira, L. A. G., Ferreira, A. M. R., and Serapião, R. V. (2018). Role of cAMP modulator supplementation during oocyte in vitro maturation on domestic animals. Reprod. Sci. 199, 1–14.
| Role of cAMP modulator supplementation during oocyte in vitro maturation on domestic animals.Crossref | GoogleScholarGoogle Scholar |
Leal, G. R., Monteiro, C. A. S., Saraiva, H. F. R. A., Camargo, A. J. R., Rodrigues, A. R. L., Oliveira, C. S., Vasconcellos, C. O. P., Nogueira, L. A. G., and Serapião, R. V. (2019). Evaluation of the simulated physiological oocyte maturation (SPOM) system on F1 Gyr × Holstein oocytes and embryos. Anim. Prod. Sci. 59, 634–640.
| Evaluation of the simulated physiological oocyte maturation (SPOM) system on F1 Gyr × Holstein oocytes and embryos.Crossref | GoogleScholarGoogle Scholar |
Li, H. J., Sutton-McDowall, M. L., Wang, X. S., Sugimura, S., Thompson, J. G., and Gilchrist, R. B. (2016). Extending prematuration with cAMP modulators enhances the cumulus contribution to oocyte antioxidant defense and oocyte quality via gap junctions. Hum. Reprod. 31, 810–821.
| Extending prematuration with cAMP modulators enhances the cumulus contribution to oocyte antioxidant defense and oocyte quality via gap junctions.Crossref | GoogleScholarGoogle Scholar | 26908844PubMed |
Luciano, A. M., Modina, S., Vassena, R., Milanesi, E., Lauria, A., and Gandolfi, F. (2004). Role of intracellular cyclic adenosine 3′,5′-monophosphate concentration and oocyte–cumulus cells communications on the acquisition of developmental competence during in vitro maturation of bovine oocytes. Biol. Reprod. 70, 465–472.
| Role of intracellular cyclic adenosine 3′,5′-monophosphate concentration and oocyte–cumulus cells communications on the acquisition of developmental competence during in vitro maturation of bovine oocytes.Crossref | GoogleScholarGoogle Scholar | 14568913PubMed |
Martino, N. A., Dell’Aquila, M. E., Filioli Uranio, M., Rutigliano, L., Nicassio, M., Lacalandra, G. M., and Hinrichs, K. (2014). Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential. Reprod. Biol. Endocrinol. 12, 99.
| Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential.Crossref | GoogleScholarGoogle Scholar | 25306508PubMed |
Metcalf, E. S., Masterson, K. R., Battaglia, D., Beck, R., and Cook, N. L. (2016). The effect of a simulated physiological oocyte maturation system on the in vitro production of equine embryos. J. Equine Vet. Sci. 41, 65.
| The effect of a simulated physiological oocyte maturation system on the in vitro production of equine embryos.Crossref | GoogleScholarGoogle Scholar |
Mottershead, D. G., Sugimura, S., Al-Musawi, S. L., Li, J. J., Richani, D., White, M. A., Martin, G. A., Trotta, A. P., Ritter, L. J., Shi, J., Mueller, T. D., Harrison, C. A., and Gilchrist, R. B. (2015). Cumulin, an oocyte-secreted heterodimer of the transforming growth factor-β family, is a potent activator of granulosa cells and improves oocyte quality. J. Biol. Chem. 290, 24007–24020.
| Cumulin, an oocyte-secreted heterodimer of the transforming growth factor-β family, is a potent activator of granulosa cells and improves oocyte quality.Crossref | GoogleScholarGoogle Scholar | 26254468PubMed |
Pedersen, H. G., Watson, E. D., and Telfer, E. E. (2004). Effect of ovary holding temperature and time on equine granulosa cell apoptosis, oocyte chromatin configuration, and cumulus morphology. Theriogenology 62, 468–480.
| Effect of ovary holding temperature and time on equine granulosa cell apoptosis, oocyte chromatin configuration, and cumulus morphology.Crossref | GoogleScholarGoogle Scholar | 15226003PubMed |
Prates, E. G., Nunes, J. T., and Pereira, R. M. (2014). A role of lipid metabolism during cumulus–oocyte complex maturation: impact of lipid modulators to improve embryo production. Mediators Inflamm. 2014, 692067.
| A role of lipid metabolism during cumulus–oocyte complex maturation: impact of lipid modulators to improve embryo production.Crossref | GoogleScholarGoogle Scholar | 24733963PubMed |
Preis, K. A., Carnevale, E. M., Coutinho da Silva, M. A., Caracciolo di Brienza, V., Gomes, G. M., Maclellan, L. J., and Squires, E. L. (2004). In vitro maturation and transfer of equine oocytes after transport of ovaries at 12 or 22 degrees C. Theriogenology 61, 1215–1223.
| In vitro maturation and transfer of equine oocytes after transport of ovaries at 12 or 22 degrees C.Crossref | GoogleScholarGoogle Scholar | 15036956PubMed |
Razza, E. M., Pedersen, H. S., Stroebech, L., Fontes, P. K., Kadarmideen, H. N., Callesen, H., Pihil, M., Nogueira, M. F. G., and Hyttel, P. (2019). Simulated physiological oocyte maturation has side effects on bovine oocytes and embryos. J. Assist. Reprod. Genet. 36, 413–424.
| Simulated physiological oocyte maturation has side effects on bovine oocytes and embryos.Crossref | GoogleScholarGoogle Scholar | 30443692PubMed |
Richani, D., Wang, X., Zeng, H. T., Smitz, J., Thompson, J. G., and Gilchrist, R. B. (2014). Pre-maturation with cAMP modulators in conjunction with EGF-like peptides during in vitro maturation enhances mouse oocyte developmental competence. Mol. Reprod. Dev. 81, 422–435.
| Pre-maturation with cAMP modulators in conjunction with EGF-like peptides during in vitro maturation enhances mouse oocyte developmental competence.Crossref | GoogleScholarGoogle Scholar | 24488930PubMed |
Richani, D., Contance, K., Lien, S., Agapioy, D., Stocker, W. A., Hedger, M. P., Ledger, W. L., Thompson, J. G., Robertson, D. M., Mottershead, D. G., Walton, K. L., Harrison, C. A., and Gilchrist, R. B. (2019). Cumulin and FSH cooperate to regulate inhibin B and activin B production by human granulosa–lutein cells in vitro. Endocrinology 160, 853–862.
| Cumulin and FSH cooperate to regulate inhibin B and activin B production by human granulosa–lutein cells in vitro.Crossref | GoogleScholarGoogle Scholar | 30753406PubMed |
Rose, R. D., Gilchrist, R. B., Kelly, J. M., Thompson, J. G., and Sutton-McDowall, M. L. (2013). Regulation of sheep oocyte maturation using cAMP modulators. Theriogenology 79, 142–148.
| Regulation of sheep oocyte maturation using cAMP modulators.Crossref | GoogleScholarGoogle Scholar | 23102843PubMed |
Russell, D. L., Gilchrist, R. B., Brown, H. M., and Thompson, J. G. (2016). Bidirectional communication between cumulus cells and the oocyte: old hands and new players? Theriogenology 86, 62–68.
| Bidirectional communication between cumulus cells and the oocyte: old hands and new players?Crossref | GoogleScholarGoogle Scholar | 27160446PubMed |
Santiquet, N. W., Greene, A. F., Becker, J., Barfield, J. P., Schoolcraft, W. B., and Krisher, R. L. (2017). A pre-in vitro maturation medium containing cumulus oocyte complex ligand–receptor signaling molecules maintains meiotic arrest, supports the cumulus oocyte complex and improves oocyte developmental competence. Mol. Hum. Reprod. 23, 594–606.
| A pre-in vitro maturation medium containing cumulus oocyte complex ligand–receptor signaling molecules maintains meiotic arrest, supports the cumulus oocyte complex and improves oocyte developmental competence.Crossref | GoogleScholarGoogle Scholar | 28586460PubMed |
Silva, G. M., Araújo, V. R., Duarte, A. B. G., Chaves, R. N., Silva, C. M. G., and Lobo, C. H. (2011). Ascorbic acid improves the survival and in vitro growth of isolated caprine preantral follicles. Anim. Reprod. 8, 14–24.
Spits, C., Guzman, L., Mertzanidou, A., Jacobs, K., Ortega-Hrepich, C., Gilchrist, R. B., Thompson, J. G., De Vos, M., Smitz, J., and Sermon, K. (2015). Chromosome constitution of human embryos generated after in vitro maturation including 3-isobutyl-1-methylxanthine in the oocyte collection medium. Hum. Reprod. 30, 653–663.
| Chromosome constitution of human embryos generated after in vitro maturation including 3-isobutyl-1-methylxanthine in the oocyte collection medium.Crossref | GoogleScholarGoogle Scholar | 25475586PubMed |
Sudiman, J., Ritter, L. J., Feil, D. K., Wang, X., Chan, K., Mottershead, D. G., Robertson, D. M., Thompson, J. G., and Gilchrist, R. B. (2014a). Effects of differing oocyte-secreted factors during mouse in vitro maturation on subsequent embryo and fetal development. J. Assist. Reprod. Genet. 31, 295–306.
| Effects of differing oocyte-secreted factors during mouse in vitro maturation on subsequent embryo and fetal development.Crossref | GoogleScholarGoogle Scholar | 24408183PubMed |
Sudiman, J., Sutton-McDowall, M. L., Ritter, L. J., White, M. A., Mottershead, D. G., Thompson, J. G., and Gilchrist, R. B. (2014b). Bone morphogenetic protein 15 in the pro-mature complex form enhances bovine oocyte developmental competence. PLoS One 9, e103563.
| Bone morphogenetic protein 15 in the pro-mature complex form enhances bovine oocyte developmental competence.Crossref | GoogleScholarGoogle Scholar | 25058588PubMed |
Torres, V., Hamdi, M., Maillo, V., Urrego, R., Echeverri, J., Lopez-Herrera, A., Gutierrez-Adan, A., Rizos, D., and Sanchez-Calabuig, M. (2019). Ascorbic acid–cyclodextrin complex alters the expression of genes associated with lipid metabolism in bovine in vitro produced embryos. Reprod. Domest. Anim. 54, 55–62.
| Ascorbic acid–cyclodextrin complex alters the expression of genes associated with lipid metabolism in bovine in vitro produced embryos.Crossref | GoogleScholarGoogle Scholar | 30120843PubMed |
Vogelsang, M. M., Kraemer, D. C., Potter, G. D., and Stott, G. G. (1987). Fine structure of the follicular oocyte of the horse. J. Reprod. Fertil. Suppl. 35, 157–167.
| 3479572PubMed |
Yu, X. X., Liu, Y. H., Liu, X. M., Wang, P. C., Liu, S., Miao, J. K., Du, Z. Q., and Yang, C. X. (2018). Ascorbic acid induces global epigenetic reprogramming to promote meiotic maturation and developmental competence of porcine oocytes. Sci. Rep. 8, 6132.
| Ascorbic acid induces global epigenetic reprogramming to promote meiotic maturation and developmental competence of porcine oocytes.Crossref | GoogleScholarGoogle Scholar | 29666467PubMed |
