Journal Search Engine
Search Advanced Search Adode Reader(link)
Download PDF Export Citaion korean bibliography PMC previewer
ISSN : 2508-755X(Print)
ISSN : 2288-0178(Online)
Journal of Embryo Transfer Vol.27 No.2 pp.71-80

The Effects of Resveratrol on Oocyte Maturation and Preimplantation Embryo Development

Sang-Hwan Hyun*, Seong-Sung Kwak
Laboratory of Veterinary Embryology and Biotechnology, College of Veterinary Medicine, Chungbuk National University


Biotechnologies for cloning animals and in vitro embryo production have the potential to produce biomedicalmodels for various researches. Especially, pigs are a suitable model for xenotransplantation, transgenic production andvarious areas of reproductive research due to its physiological similarities to human. However, utilization of in vitroproducedembryos for transfer remains limited. Despite improvement over past few decades, obstacles associated withthe production of good quality embryos in vitro still exist which limit the efficiency of cloning. One of major problemsincludes improper in vitro maturation (IVM) and culture (IVC). Oxidative stress caused from in vitro cultureconditions contributes to inadequate IVM and IVC which leads to poor developmental competence of oocytes, failureof fertilization and embryo development. To reduce the oxidative stress, various antioxidants have been used to IVMand IVC system. However, limited information is available on the effects of resveratrol on livestock reproductions.Resveratrol is a polyphenolic natural product and well known as an antioxidant in foods and beverages (e.g. in grapesand red wine). Resveratrol is known to be cardioprotective, anticarcinogenic, anti-inflammatory, antioxidant and antiapoptotic.This paper will review the effects of resveratrol on in vitro maturation of oocytes and embryo development.

01 Seong-Sung Kwak.pdf346.0KB


 Biotechnologies for producing transgenic animals and in vitro embryo production have the potential to produce biomedical animal models for various researches. Especially, pigs can be a suitable model for xenotransplantation, transgenic production and various areas of reproductive research due to its physiological similarities to human (Abeydeera, 2002). In order for the animals to remain a suitable model for research purposes, embryos which are viable and have good quality need to be produced efficiently. Recently, the in vitro culture environment is well studied and becoming more defined (Nagai, 2001; Gil et al., 2010); however, mammalian embryos produced in vitro are known to have reduced developmental competence as compared with embryos produced in vivo (Wang et al., 1997; Wang et al., 1999). Because oocytes and embryos in the in vitro culture system are vulnerable to oxidative stress. Therefore, many researches are aimed at reducing oxidative stress of this system through antioxidant treatment and elucidating the reproductive and cellular mechanisms associated with the oocytes maturation and embryos development.

 Several criteria for example the intracellular glutathione (GSH) and reactive oxygen species (ROS) levels, the maturation promoting factor activity and cortical granule exocytosis have been used as indicators to examine the cytoplasmic maturity of matured oocytes. Among them, the intracellular levels of GSH and ROS are well known to be critical factors that influence the oocyte in vitro maturation (IVM) and embryo development (Abeydeera et al., 1998; De Matos and Furnus, 2000; You et al., 2010). The ROS such as hydrogen peroxide, superoxide anions and hydroxyl radicals, are generated during intermediate steps of oxygen reduction and when ROS is overproduced and the cell cannot adapt, oxidative stress occurs. High level of ROS can damage cell membranes (Nasr-Esfahani et al., 1990), DNA (Halliwell and Aruoma, 1991), RNA transcription and protein synthesis (Takahashi et al., 2000), and might play a role in apoptosis (Yang et al., 1998). These detrimental effects of high ROS level in matured oocytes and embryos are supposed to cause detrimental effects on embryonic development and subsequently lead to early embryonic death (Guerin et al., 2001; You et al., 2010; Kwak et al., 2012).

 The intracellular GSH, a ubiquitous intracellular sulfhydryl compound, in matured oocyte is a molecular marker to predict cytoplasmic maturation in porcine oocytes (Wang et al., 1997; Abeydeera et al., 1998), and involved in various cellular processes including the synthesis of DNA and proteins, the metabolism of chemicals, cellular protection and amino acid transport (Meister and Anderson, 1983). Also, it has been found that intracellular GSH plays an important role in protecting the cell from oxidative damage (Meister, 1983) and regulates the intracellular redox metabolism (Luberda, 2005). Intracellular GSH has also been linked to increased thermo-tolerance in early bovine embryos (Edwards et al., 2001). On the other hand, low intracellular GSH concentration is responsible for lower developmental competence of porcine oocytes (Brad et al., 2003). Thus, having a high concentration of intracellular GSH and low ROS level of in vitro-matured oocytes and embryos may improve the oocytes and embryos viability and contribute to the development of assisted reproduction technologies in livestock species.

 To improve the developmental competence of oocytes in vitro matured and preimplantation embryonic development cultured in vitro, various antioxidants such as β-mercaptoethanol (β-ME), cysteine (De Matos and Furnus, 2000), cysteamine (Kobayashi et al., 2006), anthocyanin (You et al., 2010) and resveratrol (Lee et al., 2010; Kwak et al., 2012) that have been added to porcine IVM and IVC system are summarized in Table 1 and Fig. 1. Recently, it was reported that treatment of resveratrol during IVM and IVC improves developmental potential of porcine oocytes and porcine embryo development (Lee et al., 2010; Kwak et al., 2012). Resveratrol (3,4',5-trihydroxystilbene) is a naturally occurring polyphenolic product (Gusman et al., 2001; Pervaiz and Holme, 2009) phytoalexin, secondary plant metabolite, produced by the interaction of plants with microorganism found in the root of Polygonum cuspidatum, Vitis vinifera, red wines, mulberries, etc., and has the ability to protect plants against fungal and bacterial infections (Langcake et al., 1979). Resveratrol exerts various biological activities including chemopreventive, anti-inflammatory, antioxidant, antiproliferative, proapoptotic, cardioprotective and anticancer (Gusman et al., 2001; Pervaiz and Holme, 2009). More biological characteristics of resveratrol are described below.

Table 1. Various antioxidants on the porcine in vitro maturation (IVM) and in vitro culture (IVC)

 Many studies have been performed to examine the physiological functions and the biological activities of resveratrol for therapeutic purposes in humans; however, there is limited information available in livestock species oocytes and embryos on the effects of resveratrol on oocytes maturation and embryo development. In this review, therefore, we discuss the effects of resveratrol on in vitro maturation of oocytes and embryo development focused in porcine model.

1. Biology of Resveratrol

 Resveratrol is a polyphenolic natural product and has a stilbene structure (Fig. 1). Its biological function is to protect the plant under attack by pathogens such as fungi or stress conditions (Soleas et al., 1997). It exists in foods and beverages for example in grapes and red wines and is widely consumed. The effects of resveratrol are recently a hot topic of various animal and human studies as well as in embryos. Resveratrol exerts various biological activities ranging from anticancer effects to life extension. By the activation of sirtuins, a class Ⅲ of histone deacetylase with a role in lifespan determination, resveratrol increased lifespan in yeast (Saccharomyces cerevisiae) (Howitz et al., 2003), Caenorhabditis elegans, Drosophila melanogaster (Wood et al., 2004) and even in mice (Baur et al., 2006). Resveratrol is also known as SIRT1 activator (Howitz et al., 2003; Baur et al., 2006; Lagouge et al., 2006; Bai et al., 2008). However, its effects on the lifespan extension of various model organisms and its effects on SIRT1 activation remain controversial (Bass et al., 2007; Beher et al., 2009; Pacholec et al., 2010).

Fig. 1. Various antioxidants structures. All structure pictures from the Wikipedia.

 “The French paradox” is known as moderate drinking of red wine reduces the risk of heart disease (Szmitko and Verma, 2005).  Cardioprotective effects of resveratrol are related to its abilities of inhibition of vascular smooth muscle cell proliferation (Haider et al., 2005), stimulation of endolethelial nitric oxide synthase activity (Wallerath et al., 2002) and inhibition of platelet aggregation (Stef et al., 2006). In addition, it also exerts the anticancer effects that resveratrol applications prevented skin cancer development in mice (Jang et al., 1997). There have been many studies of the anticancer activity of resveratrol in cell and animal models (Jang and Surh, 2001; Baur and Sinclair, 2006). It was reported the proapoptotic activity of resveratrol in platelets (Lin et al., 2009) and smooth muscle (Zou et al., 1999). To date, however, there are no results of human clinical trials for cancer (Athar et al., 2007). Not only the above effects of resveratrol, but also it have various effects: Antidiabetic (Su et al., 2006), antiapoptotic (Jang and Surh, 2001), neuroprotective (Karuppagounder et al., 2009), anti-inflammatory (Gentilli et al., 2001) and antiviral effects (Docherty et al., 2006).

 In reproduction field, resveratrol is known to be an estrogen receptor modulator (Gehm et al., 1997; Bhat et al., 2001) and an aromatase inhibitor (Wang et al., 2006). So, resveratrol supplementation increased testosterone levels in mice (Shin et al., 2008) and it has been reported to increase sperm output in rats (Juan et al., 2005). It was reported that resveratrol, known as aryl hydrocarbon receptor antagonist, protects sperm from DNA damage and apoptosis caused by benzo (a) pyrene (Revel et al., 2001).

2. Resveratrol in In Vitro Maturation and Fertilization

 It was demonstrated that 2 μM resveratrol during IVM of porcine oocytes showed beneficial effects on oocyte maturation and subsequent embryonic development of porcine embryos derived by parthenogenetic activation (PA) and in vitro fertilization (IVF). Treatment of porcine oocytes with 2 μM resveratrol during IVM effectively reduced the intracellular level of ROS and increasing GSH concentration in mature oocytes, and subsequently reduced apoptosis-related gene expressions in matured oocytes and cumulus cells. Consequently, 2 μM resveratrol treated oocytes enhanced subsequent in vitro development of PA and IVF embryos and reduced apoptosis-related gene expression in blastocysts (Kwak et al., 2012). From this finding, resveratrol may have a role in reducing intracellular ROS in mature oocyte, and this is consistent with previous study that resveratrol is a good scavenger of ROS (Mahal and Mukherjee, 2006).

 The treatment of 10 μM resveratrol during IVM, however, significantly decreased nuclear maturation. At certain concentrations, resveratrol has been shown to exert pro-apoptotic properties in somatic cells (Kim et al., 2004) and to decrease nuclear maturation in vitro in bovine oocytes by binding to arylhydrocarbon receptors (Pocar et al., 2004). In bovine, resveratrol treatment of 20 and 40 μM during IVM significantly reduced the percentage of oocytes that reached the MⅡ stage. Therefore, high concentrations (≥ 10 μM) of resveratrol might have a detrimental effect on oocyte maturation. From the recent study (Park et al., 2012), it might be caused by the competitively inhibiting activity of various phosphodiesterases, which result in an increase in cytosolic concentration of cAMP, which can block the maturation.

 Because of the beneficial effects on cytoplasmic maturation, embryonic development and viability of blastocyst after PA and IVF were also improved. After fertilization, GSH participates in sperm decondensation in parallel to oocyte activation, and in the transformation of the fertilizing sperm head into the male pronucleus (Yoshida et al., 1992). The competence of monospermic fertilization is dependent on the degree of cytoplasmic maturation in the matured oocytes (Han et al., 1999) and subsequently influences early developmental potential (Koo et al., 2005). Taken together, it was shown that monospermic fertilization and developmental competence were greatly improved by 2 μM resveratrol treatment during IVM (Kwak et al., 2012). These results suggest that resveratrol is useful supplement on IVM medium, leading to successful cytoplasmic maturation and subsequent in vitro fertilization in porcine.

 There are some studies of the effects of resveratrol on cryopreservation and vitrification. In human, resveratrol prevents DNA damage induced by cryopreservation in human semen (Branco et al., 2010; Garcez et al., 2010). In other study, the scavenger properties of resveratrol were demonstrated in vitro in human sperm, thus resveratrol could be added to the media used in assisted reproduction techniques and cryopreservation when oxidative stress is exacerbated (Collodel et al., 2011). In ram, resveratrol treatment increase sperm motility and reduce acrosomal defect in conservation of ram semen, so it can improve the ram semen preservation (Sarlos et al., 2002). In cat, resveratrol showed reversible and beneficial effects to cat oocyte survival during vitrification (Comizzoli et al., 2009).

3. Resveratrol in Embryo Development

 It was reported that 0.5 μM resveratrol improved blastocyst formation, increased total cell numbers of PA and IVF blastocysts and affected events related to apoptosis by down-regulating the Caspase-3 mRNA expression in the developing embryos (Lee et al., 2010). This result is also in agreement with our unpublished data. In this study, high concentration (25 μM) of resveratrol in the culture medium had toxic side effects and was detrimental to the developing embryos. Similarly, in cell culture studies, resveratrol had a dose-dependent effect on the proliferation of cultured cells. At high concentrations of resveratrol induced apoptosis and decreased mitotic activity (Clement et al., 1998) whereas at low concentrations it prompted cell division in various human cell lines (Szende et al., 2000).

 In previous studies, resveratrol protected against hazardous effects of 2-bromopropane on maturation of mouse oocytes, fertilization and fetal development (Huang and Chan, 2012). In addition, resveratrol protected against methylglyoxal-induced apoptosis and disruption of embryonic development in mouse blastocysts (Huang et al., 2011a). It also reported that protective effects of resveratrol on ethanol-induced apoptosis in embryonic stem cells and disruption of embryonic development in mouse blastocysts (Huang et al., 2007).

 To date, the molecular mechanisms by which resveratrol exerts its antioxidative and antiapoptotic effects on oocyte maturation and embryonic development remains elusive. Intracellular GSH is one of the major antioxidants and plays key roles in maintaining redox homeostasis, scavenging peroxides and detoxifying xenobiotics (Meister, 1992; Hayes et al., 2005). Recent studies have shown that resveratrol could increase GSH levels (Sharma and Gupta, 2002; Savaskan et al., 2003; Yen et al., 2003; Li et al., 2006), suggesting that modulating the GSH homeostasis by resveratrol may play a significant role in the beneficial effects in oocyte maturation and embryo development.


 Resveratrol supplementation in the in vitro maturation and culture media also in the semen cryopreservation is advantageous for oocyte cytoplasmic maturation, embryo development and semen cryopreservation by reducing the intracellular ROS level, increasing GSH concentration, and down-regulation of apoptosis-related genes. Furthermore, resveratrol treatment of porcine oocytes could have better developmental competence, so greatly improve monospermic fertilization, blastocyst formation, and blastomere viability in PA and IVF-derived embryos. These findings will be useful for improving the early development of livestock embryos by reducing oxidative stress. Therefore, resveratrol treatment during IVM and IVC has the potential to increase the efficiency of cloned animal production and establishment of embryonic stem cells.


1.Abeydeera L. 2002. In vitro production of embryos in swine. Theriogenology 57:257-273.
2.Abeydeera L, Wang W, Cantley T, Prather R and Day B. 1998. Presence of [beta]-mercaptoethanol can increase the glutathione content of pig oocytes matured in vitro and the rate of blastocyst development after in vitro fertilization. Theriogenology 50:747-756.
3.Abeydeera LR, Wang WH, Cantley TC, Prather RS and Day BN. 1999. Glutathione content and embryo development after in vitro fertilisation of pig oocytes matured in the presence of a thiol compound and various concentrations of cysteine. Zygote 7:203-210.
4.Athar M, Back JH, Tang X, Kim KH, Kopelovich L, Bickers DR and Kim AL. 2007. Resveratrol: A review of preclinical studies for human cancer prevention. Toxicol. Appl. Pharmacol. 224:274-283.
5.Bai L, Pang WJ, Yang YJ and Yang GS. 2008. Modulation of Sirt1 by resveratrol and nicotinamide alters proliferation and differentiation of pig preadipocytes. Mol. Cell. Biochem. 307:129-140.
6.Bass TM, Weinkove D, Houthoofd K, Gems D and Partridge L. 2007. Effects of resveratrol on lifespan in Drosophila melanogaster and Caenorhabditis elegans. Mech. Ageing. Dev. 128:546-552.
7.Baur JA and Sinclair DA. 2006. Therapeutic potential of resveratrol: the in vivo evidence. Nat. Rev. Drug Discov. 5:493-506.
8.Baur JA, Pearson KJ, Price NL, Jamieson HA, Lerin C, Kalra A, Prabhu VV, Allard JS, Lopez-Lluch G and Lewis K. 2006. Resveratrol improves health and survival of mice on a high-calorie diet. Nature 444:337-342.
9.Beher D, Wu J, Cumine S, Kim KW, Lu SC, Atangan L and Wang M. 2009. Resveratrol is not a direct activator of SIRT1 enzyme activity. Chem. Biol. Drug Des. 74:619-624.
10.Bhat KPL, Lantvit D, Christov K, Mehta RG, Moon RC and Pezzuto JM. 2001. Estrogenic and antiestrogenic properties of resveratrol in mammary tumor models. Cancer Res. 61:7456.
11.Brad A, Bormann C, Swain J, Durkin R, Johnson A, Clifford A and Krisher R. 2003. Glutathione and adenosine triphosphate content of in vivo and in vitro matured porcine oocytes. Mol. Reprod. Dev. 64:492-498.
12.Branco CS, Garcez ME, Pasqualotto FF, Erdtman B and Salvador M. 2010. Resveratrol and ascorbic acid prevent DNA damage induced by cryopreservation in human semen. Cryobiology 60:235-237.
13.Chawalit S, Nguyen NT, Tseng JK, Lo NW, Tu CF and Ju JC. 2012. Trichostatin A and ascorbic acid assist in the development of porcine handmade cloned embryos via different physiologic pathways. Reprod. Sci. In press.
14.Choe C, Shin YW, Kim EJ, Cho SR, Kim HJ, Choi SH, Han MH, Han J, Son DS and Kang D. 2010. Synergistic effects of glutathione and β-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-ysteine. J. Reprod. Dev. 56:575-582.
15.Choi J, Park SM, Lee E, Kim JH, Jeong YI, Lee JY, Park SW, Kim HS, Hossein MS, Jeong YW, Kim S, Hyun SH and Hwang WS. 2008. Anti-apoptotic effect of melatonin on preimplantation development of porcine parthenogenetic embryos. Mol. Reprod. Dev. 75:1127-1135.
16.Clement MV, Hirpara JL, Chawdhury SH and Pervaiz S. 1998. Chemopreventive agent resveratrol, a natural product derived from grapes, triggers CD95 signaling-dependent apoptosis in human tumor cells. Blood 92:996-1002.
17.Collodel G, Federico M, Geminiani M, Martini S, Bonechi C, Rossi C, Figura N and Moretti E. 2011. Effect of trans-resveratrol on induced oxidative stress in human sperm and in rat germinal cells. Reprod. Toxicol. 31:239-246.
18.Comizzoli P, Wildt D and Pukazhenthi B. 2009. In vitro compaction of germinal vesicle chromatin is beneficial to survival of vitrified cat oocytes. Reprod. Domest. Anim. 44:269-274.
19.De Matos D and Furnus C. 2000. The importance of having high glutathione (GSH) level after bovine in vitro maturation on embryo development: Effect of [beta]-mercaptoethanol, cysteine and cystine. Theriogenology 53:761-771.
20.Docherty JJ, Sweet TJ, Bailey E, Faith SA and Booth T. 2006. Resveratrol inhibition of varicella-zoster virus replication in vitro. Antiviral Res. 72:171-177.
21.Edwards J, King W, Kawarsky S and Ealy A. 2001. Responsiveness of early embryos to environmental insults: potential protective roles of HSP70 and glutathione. Theriogenology 55:209-223.
22.Gajda B, Bryla M and Smorag Z. 2008. Effects of protein source, vitamin E and phenazine ethosulfate on developmental competence and quality of porcine embryos cultured in vitro. Folia Biol. 56:57-63.
23.Garcez ME, dos Santos Branco C, Lara LV, Pasqualotto FF and Salvador M. 2010. Effects of resveratrol supplementation on cryopreservation medium of human semen. Fertil. Steril. 94:2118-2121.
24.Gehm BD, McAndrews JM, Chien PY and Jameson JL. 1997. Resveratrol, a polyphenolic compound found in grapes and wine, is an agonist for the estrogen receptor. Proc. Natl. Acad. Sci. USA 94:14138-14143.
25.Gentilli M, Mazoit JX, Bouaziz H, Fletcher D, Casper RF, Benhamou D and Savouret JF. 2001. Resveratrol decreases hyperalgesia induced by carrageenan in the rat hind paw. Life Sci. 68:1317-1321.
26.Gil M, Cuello C, Parrilla I, Vazquez J, Roca J and Martinez E. 2010. Advances in swine in vitro embryo production technologies. Reprod. Domest. Anim. 45:40-48.
27.Grupen C, Nagashima H and Nottle M. 1995. Cysteamine enhances in vitro development of porcine oocytes matured and fertilized in vitro. Biol. Reprod. 53:173-178.
28.Guerin P, El Mouatassim S and Menezo Y. 2001. Oxidative stress and protection against reactive oxygen species in the pre-implantation embryo and its surroundings. Hum. Reprod. Update 7: 175-189.
29.Gusman J, Malonne H and Atassi G. 2001. A reappraisal of the potential chemopreventive and chemotherapeutic properties of resveratrol. Carcinogenesis 22:1111-1117.
30.Haider UGB, Roos TU, Kontaridis MI, Neel BG, Sorescu D, Griendling KK, Vollmar AM and Dirsch VM. 2005. Resveratrol inhibits angiotensin Ⅱ-and epidermal growth factormediated Akt activation: role of Gab1 and Shp2. Mol. Pharmacol. 68:41-48.
31.Halliwell B and Aruoma OI. 1991. DNA damage by oxygen-derived species Its mechanism and measurement in mammalian systems. FEBS letters 281:9-19.
32.Han YM, Wang WH, Abeydeera LR, Petersen AL, Kim JH, Murphy C, Day BN and Prather RS. 1999. Pronuclear location before the first cell division determines ploidy of polyspermic pig embryos. Biol. Reprod. 61:1340-1346.
33.Hayes JD, Flanagan JU and Jowsey IR. 2005. Glutathione transferases. Annu. Rev. Pharmacol. Toxicol. 45:51-88.
34.Hossein MS, Hashem MA, Jeong YW, Lee MS, Kim S, Kim JH, Koo OJ, Park SM, Lee EG, Park SW, Kang SK, Lee BC and Hwang WS. 2007. Temporal effects of a-tocopherol and l-ascorbic acid on in vitro fertilized porcine embryo development. Anim. Reprod. Sci. 100:107-117.
35.Howitz KT, Bitterman KJ, Cohen HY, Lamming DW, Lavu S, Wood JG, Zipkin RE, Chung P, Kisielewski A and Zhang LL. 2003. Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature 425: 191-196.
36.Hu J, Cheng D, Gao X, Bao J, Ma X and Wang H. 2012. Vitamin C enhances the in vitro development of porcine preimplantation embryos by reducing oxidative stress. Reprod. Domest. Anim. In press.
37.Huang FJ, Chin TY and Chan WH. 2011a. Resveratrol protects against methylglyoxal-induced apoptosis and disruption of embryonic development in mouse blastocysts. Environ. Toxicol. In press.
38.Huang IM and Chan WH. 2012. Resveratrol protects against hazardous effects of 2-bromopropane on maturation of mouse oocytes, fertilization and fetal development. Afr. J. Biotechnol. 11:10262-10271.
39.Huang LH, Shiao NH, Hsuuw YD and Chan WH. 2007. Protective effects of resveratrol on ethanol-induced apoptosis in embryonic stem cells and disruption of embryonic development in mouse blastocysts. Toxicology 242:109-122.
40.Huang Y, Tang X, Xie W, Zhou Y, Li D, Zhu J, Yuan T, Lai L and Pang D. 2011b. Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos. Biochem. Biophys. Res. Commu. 411:397-401.
41.Jang JH and Surh YJ. 2001. Protective effects of resveratrol on hydrogen peroxide-induced apoptosis in rat pheochromocytoma (PC12) cells. Mutat. Res.-Genet. Toxicol. Environ. Mutat. Res. 496:181-190.
42.Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CWW, Fong HHS, Farnsworth NR, Kinghorn AD and Mehta RG. 1997. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275:218-220.
43.Jeong BS and Yang X. 2001. Cysteine, glutathione, and percoll treatments improve porcine oocyte maturation and fertilization in vitro. Mol. Reprod. Dev. 59:330-335.
44.Jeong YW, Hossein MS, Bhandari DP, Kim YW, Kim JH, Park SW, Lee E, Park SM, Jeong YI, Lee JY, Kim S and Hwang WS. 2008. Effects of insulin-transferrin-selenium in defined and porcine follicular fluid supplemented IVM media on porcine IVF and SCNT embryo production. Anim. Reprod. Sci. 106:13-24.
45.Jeong YW, Park SW, Hossein MS, Kim S, Kim JH, Lee SH, Kang SK, Lee BC and Hwang WS. 2006. Antiapoptotic and embryotrophic effects of a-tocopherol and L-ascorbic acid on porcine embryos derived from in vitro fertilization and somatic cell nuclear transfer. Theriogenology 66:2104-2112.
46.Juan ME, Gonzalez-Pons E, Munuera T, Ballester J, Rodriguez-Gil JE and Planas JM. 2005. trans-Resveratrol, a natural antioxidant from grapes, increases sperm output in healthy rats. J. Nutr. 135:757-760.
47.Kang JT, Koo OJ, Kwon DK, Park HJ, Jang G, Kang SK and Lee BC. 2009. Effects of melatonin on in vitro maturation of porcine oocyte and expression of melatonin receptor RNA in cumulus and granulosa cells. J. Pineal Res. 46:22-28.
48.Karuppagounder SS, Pinto JT, Xu H, Chen HL, Beal MF and Gibson GE. 2009. Dietary supplementation with resveratrol reduces plaque pathology in a transgenic model of Alzheimer's disease. Neurochem. Int. 54:111-118.
49.Kim Y, Byung T, Yong TL, Rhee SH and Park KY. 2004. Resveratrol inhibits cell proliferation and induces apoptosis of human breast carcinoma MCF-7 cells. Oncol. Rep. 11:441-446.
50.Kitagawa Y, Suzuki K, Yoneda A and Watanabe T. 2004. Effects of oxygen concentration and antioxidants on the in vitro developmental ability, production of reactive oxygen species (ROS), and DNA fragmentation in porcine embryos. Theriogenology 62:1186-1197.
51.Kobayashi M, Lee ES and Fukui Y. 2006. Cysteamine or [beta]-mercaptoethanol added to a defined maturation medium improves blastocyst formation of porcine oocytes after intracytoplasmic sperm injection. Theriogenology 65:1191-1199.
52.Koo DB, Kim YJ, Yu I, Kim HN, Lee KK and Han YM. 2005. Effects of in vitro fertilization conditions on preimplantation development and quality of pig embryos. Anim. Reprod. Sci. 90: 101-110.
53.Kwak SS, Cheong SA, Jeon Y, Lee E, Choi KC, Jeung EB and Hyun SH. 2012. The effects of resveratrol on porcine oocyte in vitro maturation and subsequent embryonic development after parthenogenetic activation and in vitro fertilization. Theriogenology 78:86-101.
54.Lagouge M, Argmann C, Gerhart-Hines Z, Meziane H, Lerin C, Daussin F, Messadeq N, Milne J, Lambert P and Elliott P. 2006. Resveratrol improves mitochondrial function and protects against metabolic disease by activating SIRT1 and PGC-1 [alpha]. Cell 127:1109-1122.
55.Langcake P, Cornford C and Pryce R. 1979. Identification of pterostilbene as a phytoalexin from Vitis vinifera leaves. Phytochemistry 18:1025-1027.
56.Lee K, Wang C, Chaille JM and Machaty Z. 2010. Effect of resveratrol on the development of porcine embryos produced in vitro. J. Reprod. Dev. 56:330-335.
57.Li Y, Cao Z and Zhu H. 2006. Upregulation of endogenous antioxidants and phase 2 enzymes by the red wine polyphenol, resveratrol in cultured aortic smooth muscle cells leads to cytoprotection against oxidative and electrophilic stress. Pharmacol. Res. 53:6-15.
58.Lin KH, Hsiao G, Shih CM, Chou DS and Sheu JR. 2009. Mechanisms of resveratrol-induced platelet apoptosis. Cardiovasc. Res. 83:575-585.
59.Luberda Z. 2005. The role of glutathione in mammalian gametes. Reprod Biol 5:5-17.
60.Mahal H and Mukherjee T. 2006. Scavenging of reactive oxygen radicals by resveratrol: antioxidant effect. Res. Chem. Intermed. 32:59-71.
61.Meister A and Anderson ME. 1983. Glutathione. Annu. Rev. Biochem. 52:711-760.
62.Meister A. 1983. Selective modification of glutathione metabolism. Science 220:472.
63.Meister A. 1992. Biosynthesis and functions of glutathione, an essential biofactor. J. Nutr. Sci. Vitaminol. 1-6.
64.Miclea I, Hettig A, Zahan M, Roman I and Miclea V. 2009. The effect of several a-tocopherol concentrations on swine oocyte maturation and embryo culture. Bulletin UASVM Ani. Sci. Biotechnol. 66:1-2.
65.Nagai T. 2001. The improvement of in vitro maturation systems for bovine and porcine oocytes. Theriogenology 55:1291-1301.
66.Nasr-Esfahani MH, Aitken JR and Johnson MH. 1990. Hydrogen peroxide levels in mouse oocytes and early cleavage stage embryos developed in vitro or in vivo. Development 109: 501-507.
67.Ozawa M, Nagai T, Fahrudin M, Karja NWK, Kaneko H, Noguchi J, Ohnuma K and Kikuchi K. 2006. Addition of glutathione or thioredoxin to culture medium reduces intracellular redox status of porcine IVM/IVF embryos, resulting in improved development to the blastocyst stage. Mol. Reprod. Dev. 73:998-1007.
68.Pacholec M, Bleasdale JE, Chrunyk B, Cunningham D, Flynn D, Garofalo RS, Griffith D, Griffor M, Loulakis P and Pabst B. 2010. SRT1720, SRT2183, SRT1460, and resveratrol are not direct activators of SIRT1. J. Biol. Chem. 285:8340-8351.
69.Park C, Lee J, Cheong H, Yang B and Kim C. 1997. Effect of superoxide dismutase (SOD) on pronucleus formation of porcine oocytes fertilized in vitro. Theriogenology 48:1137-1146.
70.Park SJ, Ahmad F, Philp A, Baar K, Williams T, Luo H, Ke H, Rehmann H, Taussig R and Brown AL. 2012. Resveratrol Ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases. Cell 148:421-433.
71.Pervaiz S and Holme AL. 2009. Resveratrol: Its biologic targets and functional activity. Antioxidants & Redox Signaling 11:2851-2897.
72.Pocar P, Augustin R and Fischer B. 2004. Constitutive expression of CYP1A1 in bovine cumulus oocyte-complexes in vitro: mechanisms and biological implications. Endocrinology 145: 1594-1601.
73.Revel A, Raanani H, Younglai E, Xu J, Han R, Savouret JF and Casper RF. 2001. Resveratrol, a natural aryl hydrocarbon receptor antagonist, protects sperm from DNA damage and apoptosis caused by benzo (a) pyrene. Reprod. Toxicol. 15: 479-486.
74.Rodriguez-Osorio N, Kim I, Wang H, Kaya A and Memili E. 2007. Melatonin increases cleavage rate of porcine preimplantation embryos in vitro. J. Pineal Res. 43:283-288.
75.Sarlos P, Molnar A and Kokai M. 2002. Comparative evaluation of the effect of antioxidants in the conservation of ram semen. Acta Veterinaria Hungarica 50:235-245.
76.Savaskan E, Olivieri G, Meier F, Seifritz E, Wirz-Justice A and Muller-Spahn F. 2003. Red wine ingredient resveratrol protects from β-amyloid neurotoxicity. Gerontology 49:380-383.
77.Sharma M and Gupta Y. 2002. Chronic treatment with trans resveratrol prevents intracerebroventricular streptozotocin induced cognitive impairment and oxidative stress in rats. Life Sci. 71:2489-2498.
78.Shi JM, Tian XZ, Zhou GB, Wang L, Gao C, Zhu SE, Zeng SM, Tian JH and Liu GS. 2009. Melatonin exists in porcine follicular fluid and improves in vitro maturation and parthenogenetic development of porcine oocytes. J. Pineal Res. 47:318-323.
79.Shin S, Jeon JH, Park D, Jang MJ, Choi JH, Choi BH, Joo SS, Nahm SS, Kim JC and Kim YB. 2008. Trans-Resveratrol relaxes the corpus cavernosum ex vivo and enhances testosterone levels and sperm quality in vivo. Arch. Pharm. Res. 31:83-87.
80.Soleas GJ, Diamandis EP and Goldberg DM. 1997. Resveratrol: A molecule whose time has come? And gone? Clin. Biochem. 30:91-113.
81.Spinaci M, Volpe S, De Ambrogi M, Tamanini C and Galeati G. 2008. Effects of epigallocatechin-3-gallate (EGCG) on in vitro maturation and fertilization of porcine oocytes. Theriogenology 69:877-885.
82.Stef G, Csiszar A, Lerea K, Ungvari Z and Veress G. 2006. Resveratrol inhibits aggregation of platelets from high-risk cardiac patients with aspirin resistance. J. Cardiovas. Pharmacol. 48: 1-5.
83.Su HC, Hung LM and Chen JK. 2006. Resveratrol, a red wine antioxidant, possesses an insulin-like effect in streptozotocin-induced diabetic rats. Am. J. Physiol. Endocrinol. Metab. 290:E1339-E1346.
84.Szende B, Tyihak E and Kiraly-Veghely Z. 2000. Dose-dependent effect of resveratrol on proliferation and apoptosis in endothelial and tumor cell cultures. Exp. Mol. Med. 32:88-92.
85.Szmitko PE and Verma S. 2005. Red wine and your heart. Circulation 111:e10-e11.
86.Takahashi M, Keicho K, Takahashi H, Ogawa H, Schulte R and Okano A. 2000. Effect of oxidative stress on development and DNA damage in in-vitro cultured bovine embryos by comet assay. Theriogenology 54:137-145.
87.Tao Y, Chen H, Tian N, Huo D, Li G, Zhang Y, Liu Y, Fang F, Ding J and Zhang X. 2010. Effects of l-ascorbic acid, a-tocopherol and co-culture on in vitro developmental potential of porcine cumulus cells free oocytes. Reprod. Domest. Anim. 45:19-25.
88.Tao Y, Zhou B, Xia G, Wang F, Wu Z and Fu M. 2004. Exposure to l-ascorbic acid or a-tocopherol facilitates the development of porcine denuded oocytes from metaphase Ⅰ to metaphase Ⅱ and prevents cumulus cells from fragmentation. Reprod. Domest. Anim. 39:52-57.
89.Tatemoto H, Okuda T, Sogo N and Muto N. 2001a. Male pronuclear formation and blastocyst formation are improved by supplementation of ascorbic acid 2-O-a-glucoside during in vitro maturation culture of denuded porcine oocytes. J. Reprod. Dev. 47:329-339.
90.Tatemoto H, Ootaki K, Shigeta K and Muto N. 2001b. Enhancement of developmental competence after in vitro fertilization of porcine oocytes by treatment with ascorbic acid 2-O-a-glucoside during in vitro maturation. Biol. Reprod. 65:1800-1806.
91.Uhm SJ, Gupta MK, Das ZC, Yang JH and Lee HT. 2010. Effect of 3-hydroxyflavone on pig embryos produced by parthenogenesis or somatic cell nuclear transfer. Reprod. Toxicol. 31:231-238.
92.Uhm SJ, Gupta MK, Yang JH, Lee SH and Lee HT. 2007. Selenium improves the developmental ability and reduces the apoptosis in porcine parthenotes. Mol. Reprod. Dev. 74:1386-1394.
93.Viet Linh N, Dang-Nguyen TQ, Nguyen BX, Manabe N and Nagai T. 2009. Effects of cysteine during in vitro maturation of porcine oocytes under low oxygen tension on their subsequent in vitro fertilization and development. J. Reprod. Dev. 55:594-598.
94.Wallerath T, Deckert G, Ternes T, Anderson H, Li H, Witte K and Forstermann U. 2002. Resveratrol, a polyphenolic phytoalexin present in red wine, enhances expression and activity of endothelial nitric oxide synthase. Circulation 106:1652-1658.
95.Wang W, Abeydeera L, Cantley T and Day B. 1997. Effects of oocyte maturation media on development of pig embryos produced by in vitro fertilization. Reproduction 111:101-108.
96.Wang WH and Day BN. 2002. Development of porcine embryos produced by IVM/IVF in a medium with or without protein supplementation: effects of extracellular glutathione. Zygote 10:109-115.
97.Wang WH, Abeydeera LR, Han YM, Prather RS and Day BN. 1999. Morphologic evaluation and actin filament distribution in porcine embryos produced in vitro and in vivo. Biol. Reprod. 60: 1020-1028.
98.Wang Y, Lee KW, Chan FL, Chen S and Leung LK. 2006. The red wine polyphenol resveratrol displays bilevel inhibition on aromatase in breast cancer cells. Toxicol. Sci. 92:71-77.
99.Watanabe H, Okawara S, Bhuiyan M and Fukui Y. 2010. Effect of lycopene on cytoplasmic maturation of porcine oocytes in vitro. Reprod. Domest. Anim. 45:838-845.
100.Whitaker B and Knight J. 2010. Effects of N-acetyl-cysteine and N-acetyl-cysteine-amide supplementation on in vitro matured porcine oocytes. Reprod. Domest. Anim. 45:755-759.
101.Wood JG, Rogina B, Lavu S, Howitz K, Helfand SL, Tatar M and Sinclair D. 2004. Sirtuin activators mimic caloric restriction and delay ageing in metazoans. Nature 430:686-689.
102.Yang HW, Hwang KJ, Kwon HC, Kim HS, Choi KW and Oh KS. 1998. Detection of reactive oxygen species (ROS) and apoptosis in human fragmented embryos. Hum. Reprod. 13:998-1002.
103.Yen GC, Duh PD and Lin CW. 2003. Effects of resveratrol and 4-hexylresorcinol on hydrogen peroxide-induced oxidative DNA damage in human lymphocytes. Free Radical Research 37:509-514.
104.Yoshida M, Ishigaki K and Pursel VG. 1992. Effect of maturation media on male pronucleus formation in pig oocytes matured in vitro. Mol. Reprod. Dev. 31:68-71.
105.You J, Kim J, Lim J and Lee E. 2010. Anthocyanin stimulates in vitro development of cloned pig embryos by increasing the intracellular glutathione level and inhibiting reactive oxygen species. Theriogenology 74:777-785.
106.Yuh HS, Yu DH, Shin MJ, Kim HJ, Bae KB, Lee DS, Lee HC, Chang WK, Park SB and Lee SG. 2010. The effects of various antioxidants on the development of parthenogenetic porcine embryos. In Vitro Cell. Dev. Biol. Anim. 46:148-154.
107.Zou J, Huang Y, Chen Q, Wang N, Cao K, Hsieh T and Wu J. 1999. Suppression of mitogenesis and regulation of cell cycle traverse by resveratrol in cultured smooth muscle cells. Int. J. Oncol. 15:647-651.