Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Shi, Wenqing; Zhu, Shi-En; Zhang, Dong; Wang, Weihua; Tang, Guoliang; Hou, Yunpeng; Tian, Shujun

doi:10.1530/rep.1.00899

Cited by 74 publications

(63 citation statements)

References 45 publications

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“…Since high lipid content makes the cytoplasm of porcine oocytes dark under observation with transillumination, the fluorescent FDA probe seems to be the fastest and most reliable staining method to separate live and dead oocytes. Also, FDA test does not seem to compromise the developmental competence of porcine oocytes [20]. Selection of live oocytes by simple observation of morphology (shape, color and lipid granulation) under a stereomicroscope is possible as well; it can be advantageous as being a rather time-saving method.…”

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confidence: 99%

Factors Affecting Cryopreservation of Porcine Oocytes

Somfai

Kikuchi

Nagai

2012

Journal of Reproduction and Development

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confidence: 99%

Factors Affecting Cryopreservation of Porcine Oocytes

Somfai

Kikuchi

Nagai

2012

Journal of Reproduction and Development

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“…It has been reported that the normality of the spindles of MII-stage oocytes immediately after freezing can be improved by paclitaxel treatment in both pigs [16] and cattle [17]. Furthermore, the developmental potency of cryopreserved MII-stage oocytes has been shown to be improved by paclitaxel treatment in mice [18], cattle [19], sheep [20] and pigs [16]. The aim of the present study was to investigate whether a combination of delipation and paclitaxel treatment would lead to efficient cryopreservation of porcine MII-stage oocytes.…”

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confidence: 99%

Developmental Ability of Porcine In Vitro Matured Oocytes at the Meiosis II Stage After Vitrification

Ogawa

Ueno

Nakayama

et al. 2010

Journal of Reproduction and Development

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Abstract. The aim of the present study was to investigate whether a combination of cytoplasmic lipid removal (delipation) and treatment by a microtubule stabilizer, paclitaxel, would lead to efficient cryopreservation of porcine in vitro matured (IVM) oocytes at the meiosis II (MII) stage. Vitrification and subsequent re-warming and culture of 109 untreated oocytes produced only 9 blastocysts (8.3%). On the other hand, the post-vitrification blastocyst rate was significantly improved (21/113, 18.6%, P<0.05) when oocytes were treated with 1 μM paclitaxel. Oocyte delipation also significantly increased the post-vitrification blastocyst rate compared with the untreated group (15/37, 40.5%, P<0.05). The delipation-and-paclitaxel group exhibited a significantly higher blastocyst rate (34/75, 45.3%, P<0.05) than the paclitaxel group, although it was not significantly higher than that for the delipation group. In transfer experiment, a total of 109 (18.6%) parthenogenetic blastocysts were obtained from 586 oocytes vitrified with the delipation-andpaclitaxel treatment. Transfer of 72 blastocysts to two recipients resulted in 14 (19.4%) somite stage fetuses. In conclusion, we demonstrated for the first time that by removing cytoplasmic lipid droplets from oocytes and performing a microtubule stabilization procedure, vitrified porcine IVM MII-stage oocytes could efficiently develop to the blastocyst stage while retaining the ability to develop into fetuses. Key words: Cryopreservation, Delipation, In vitro matured oocytes, Pig, Vitrification (J. Reprod. Dev. 56: [356][357][358][359][360][361] 2010) ryopreservation techniques for mammalian oocytes and embryos have been used for three main purposes: to preserve the genes of elite livestock animals and increase the efficiency of animal breeding, preserve valuable genetically modified animals and endangered species and use germ cells effectively in assisted reproductive technology. Thus far, embryo cryopreservation as a practical technique has been used in a variety of species, including experimental and livestock animals [1,2]. On the other hand, cryopreservation of oocytes remains impractical compared with embryos, and successful production of progeny from cryopreserved oocytes has been reported in limited species, including rabbits [3] [8] and rats [9]. As with porcine embryos, unfertilized porcine oocytes are highly sensitive to low temperature [10]. Consequently, cryopreserved porcine oocytes have yet to be used to successfully produce viable piglets.One of the reasons for the low-temperature sensitivity of porcine oocytes and embryos is their high intracellular lipid content. Nagashima and colleagues showed that the freezing tolerance of porcine oocytes [11] and early-stage embryos [12,13] can be dramatically increased by removing their cytoplasmic lipid droplets using a process called delipation.It is known that the meiotic spindle of the oocyte is extremely cryosensitive, and the impaired development of oocytes at meiosis II after cryopreservation has been attribut...

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“…However, it is well known that the vitrification procedure exerts a detrimental effect(s) on oocytes; for example, the most serious damage after vitrification is disorganization of spindles potentially caused by microtubule disassembly in vitrified oocytes [40]. Recently, paclitaxel treatment prior to vitrification has been shown to improve the developmental ability of mouse [41], human [42], bovine [43] and porcine [44] oocytes. Paclitaxel is a chemical that promotes formation of highly stable microtubules that resist depolymerization, thus preventing normal cell division and arresting the cell cycle at the M-phase in somatic cells [45,46].…”

Section: Discussionmentioning

confidence: 99%

Ethylene Glycol-supplemented Calcium-free Media Improve Zona Penetration of Vitrified Rat Oocytes by Sperm Cells

Fujiwara

Sano

Seita

et al. 2010

Journal of Reproduction and Development

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Abstract. Cryopreservation of matured oocytes is a useful technique because the oocytes can be used for some assisted reproductive technologies after warming. Even though rats, like mice, have been used in various research fields including reproductive technology, information about cryopreservation of rat oocytes is limited. The objective of the present study was to improve the vitrification protocol for matured rat oocytes. To determine the optimal equilibration time, oocytes were equilibrated in 7.5% ethylene glycol (EG) + 7.5% dimethyl sulfoxide (DMSO) + 20% fetal calf serum (FCS) for 1, 4, 7 or 10 min at 24 C and then 15% EG + 15% DMSO + 0.5 M sucrose + 20% FCS for 1 min at 24 C before being plunged into liquid nitrogen on Cryotops. Oocytes exposed to equilibration medium for 4 min showed higher survival and cleavage rates after artificial activation than those of oocytes exposed for 1, 7 or 10 min. The survival and cleavage rates of vitrified oocytes after activation were 98.3 and 78.4%, respectively. However, the perivitelline spaces of most of the vitrified/warmed oocytes (6/168, 3.6%) could not be penetrated by sperm after in vitro fertilization, and cortical granule exocytosis (CGE) was observed in the oocytes. Therefore, the inhibitory effect of calcium and cryoprotectants in vitrification medium on CGE was examined. In most of the oocytes vitrified in calcium-free media, CGE was strongly suppressed independent of cryoprotectants. Oocytes vitrified in EG-supplemented calcium-free media showed high survival rates after warming (79.4%). After artificial activation, the cleavage and blastocyst formation rates of the oocytes were also high (72.8 and 23.1%, respectively). The zona penetration rate of vitrified/ warmed oocytes was dramatically improved by using EG-supplemented calcium-free media after in vitro fertilization (111/155, 63.9%). Thus, our data suggest that EG-supplemented calcium-free media improve zona penetration of vitrified rat oocytes by sperm cells. Key words: Cryopreservation, Oocytes, Rat, Vitrification (J. Reprod. Dev. 56: [169][170][171][172][173][174][175] 2010) ryopreservation of germ cells and reproductive organs is a useful and important technology for efficient production and maintenance of transgenic, mutant, knock-out and other animals. In particular, embryo and sperm cryopreservation has been routinely used for not only efficient production of experimental animals but also clinical medicine. On the other hand, the most desired germ cells for freezing or vitrification are matured oocytes because successfully cryopreserved oocytes can be used for in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) or somatic-cell nuclear transfer (SCNT) [1,2]. Although successful freezing or vitrification of oocytes has been reported in several mammalian species [3][4][5][6], the survival is generally low. Especially in rats, a high rate of successful vitrification of early-stage embryos has not been obtained until recent years, possibly because rat germ cells seem to be sens...

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Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Cited by 74 publications

References 45 publications

Factors Affecting Cryopreservation of Porcine Oocytes

Factors Affecting Cryopreservation of Porcine Oocytes

Developmental Ability of Porcine In Vitro Matured Oocytes at the Meiosis II Stage After Vitrification

Ethylene Glycol-supplemented Calcium-free Media Improve Zona Penetration of Vitrified Rat Oocytes by Sperm Cells

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