Developmental potential of bovine oocytes cultured in different maturation and culture conditions

Sağırkaya, Hakan; Mısırlıoğlu, Müge; Kaya, Abdullah; First, N. L.; Parrish, J.J.; Memili, Erdoğan

doi:10.1016/j.anireprosci.2006.09.016

Cited by 73 publications

(55 citation statements)

References 40 publications

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“…The optimal conditions determined in this study included a shorter exposure time and lower concentration of TSA compared with the previous studies [16,31]. This difference might be due to the difference in nuclear transfer protocol and/or culture medium because previous studies have reported that the nuclear transfer protocol [32] and culture medium [33] affect the gene expression of embryos, including DNA methyltransferases, which are related to epigenetic modification. Thus, a difference in expression of epigenetic modification related genes might affect the histone acetylation status in NT embryos, causing a difference in the optimal treatment.…”

mentioning

confidence: 81%

Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos

Yamanaka

Sugimura

Wakai³

et al. 2009

J. Reprod. Dev.

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Abstract.Successful cloning by somatic cell nuclear transfer (SCNT) requires a reprogramming process in which the epigenetic state of a differentiated donor nucleus must be converted into an embryonic totipotent state. However, this epigenetic reprogramming is incomplete in SCNT embryos, causing low production efficiency. Recently, it has been reported that trichostatin A (TSA), an inhibitor of histone deacetylase, potentially enhances cloning efficiency. The aim of the present study was to optimize the TSA treatment for miniature pig SCNT embryos and investigate the effect of the acetylation level of histone on developmental competence of SCNT embryos. In order to optimize the TSA treatment, we examined the developmental competence of SCNT embryos under various exposure times (0-50 h) and concentrations (0-500 nM). Treatment with 5 nM TSA for 15 and 20 h beginning at the start of activation significantly increased the blastocyst formation rate (34.6 and 32.4 vs. 18.2%, respectively) and mean cell number (57.0 ± 2.7 and 56.6 ± 2.7 vs. 43.5 ± 2.1, respectively) as compared with the non-treated group (0 h). We then investigated the acetylation levels of histone H3 in SCNT embryos treated with or without TSA (TSA (+) or TSA (-)) as compared with in vitrofertilized (IVF) embryos. The acetylation levels of the TSA (-) SCNT embryos at the pseudo-pronuclear and 2-cell stages were significantly lower than those of the IVF embryos at the same developmental stages. In contrast, the acetylation levels of the TSA (+) SCNT embryos were similar to those of the IVF embryos. There was no difference in the acetylation levels of all groups at the blastocyst stage. Our data therefore suggests that the acetylation level of histone H3 at the pseudo-pronuclear and 2-cell stages is positively correlated with subsequent development of SCNT embryos, which may be an important event for the vital development of SCNT embryos in miniature pigs. Key words: Embryo development, Histone acetylation, Miniature pig, Nuclear transfer, Trichostatin A (J. Reprod. Dev. 55: [638][639][640][641][642][643][644] 2009) lthough many studies have been performed to improve the developmental competence of miniature pig SCNT embryos [1][2][3][4], the overall efficiency remains low. To obtain sufficient developmental competence for SCNT embryos to develop to term, the differentiated cell nucleus should be subjected to epigenetic reprogramming processes including chromatin remodeling and DNA methylation during preimplantation development. However, reprogramming of a differentiated nucleus to an embryonic state is reportedly delayed and incomplete in SCNT embryos [5]. In fact, previous studies have demonstrated that epigenetic modifications such as DNA methylation and histone acetylation are abnormally reprogrammed in SCNT embryos during early development [6,7]. Incomplete reprogramming of epigenetic modifications causes abnormal gene expression including imprinted genes [8], resulting in deleterious effects on the development of SCNT embryos [9].Histone acety...

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mentioning

confidence: 81%

Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos

Yamanaka

Sugimura

Wakai³

et al. 2009

J. Reprod. Dev.

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“…This issue is of particular relevance to IVG and maturation strategies as epigenetic information that is essential for normal development is acquired during gametogenesis and a growing body of evidence suggests that assisted reproductive technologies, and especially those involving extended culture and invasive micromanipulations may affect these complex processes and lead to disorders of genomic imprinting (Huntriss & Picton 2008). Imprinted genes including the maternally expressed H19 gene, IGF2 and also epigenetic regulators such as members of the DNA methyltransferases family DNMT1 and DNMT3A have been shown to be aberrantly expressed following assisted reproduction technologies (FernandezGonzalez et al 2004, Sagirkaya et al 2007). Analysis of the DNA methylation status of differentially methylated regions of the imprinted genes H19, paternally expressed gene 1/mesoderm-specific transcript (Peg1/Mest) and Igf2r in fully grown murine, germinal vesicle-staged oocytes derived in vitro from preantral follicles compared with in vivo grown counterparts indicated a loss of methylation at the Igf2r and Mest/Peg1 loci, as well as a gain of methylation of H19 (Kerjean et al 2003).…”

Section: The Follicle Culture Environment In Vitromentioning

confidence: 99%

The in vitro growth and maturation of follicles

Picton

Harris²,

Muruvi³

et al. 2008

Reproduction

242

190

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The development of technologies to grow oocytes from the most abundant primordial follicles to maturity in vitro holds many attractions for clinical practice, animal production technology and research. The production of fertile oocytes and live offspring has been achieved in mice following the long-term culture of oocytes in primordial follicles from both fresh and cryopreserved ovarian tissue. In contrast, in non-rodent species advances in follicle culture are centred on the growth of isolated preantral follicles. As a functional unit, mammalian preantral follicles are well-suited to culture but primordial and primary follicles do not grow well after isolation from the ovarian stroma. The current challenges for follicle culture are numerous and include: optimisation of culture media and the tailoring of culture environments to match the physiological needs of the cell in vivo; the maintenance of cell-cell communication and signalling during culture; and the evaluation of the epigenetic status, genetic health and fertility of in vitro derived mature oocytes. In large animals and humans, the complete in vitro growth and maturation of oocytes is only likely to be achieved following the development of a multistage strategy that closely mimics the ovary in vivo. In this approach, primordial follicle growth will be initiated in situ by the culture of ovarian cortex. Isolated preantral follicles will then be grown to antral stages before steroidogenic function is induced in the somatic cells. Finally, cytoplasmic and nuclear maturation will be induced in the in vitro derived oocytes with the production of fertile metaphase II gametes.

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“…Ovarium sapi yang berasal dari rumah potong hewan (RPH) sesaat setelah penyembelihan dapat dimanfaatkan sebagai sumber oosit untuk keperluan in vitro maturasi sehingga dapat memudahkan in vitro fertilisasi (Pujol et al, 2004), namun keberhasilan IVF sampai ke tahap blastosist sangat tergantung pada beberapa faktor diantaranya jenis suplemen yang digunakan dalam media maturasi in vitro (Hammam et al, 2010), kualitas oosit yang digunakan (Lonergan et al, 2003;Anguita et al, 2007), serta resiko kontaminasi dan kondisi kultur (Sagirkaya et al, 2007). Jenis suplemen, kualitas oosit serta kondisi kultur yang baik sangat mendukung untuk meningkatkan kemampuan maturasi oosit in vitro.…”

Section: Pendahuluanunclassified

Pengaruh Suplementasi Fetal Calf Serum Terhadap Kemampuan Maturasi in Vitro Oosit Sapi

Daoed¹,

Ngadiyono²,

Widayati³

2014

BuletinPeternak

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INTISARIPenelitian ini memanfaatkan hasil samping rumah potong hewan (RPH) sebagai sumber oosit untuk in vitro fertilization (IVF). Untuk meningkatkan keberhasilan IVF dilakukan suplementasi fetal calf serum (FCS) pada medium maturasi in vitro. Ovarium sapi dari RPH dibawa ke laboratorium dalam medium NaCl 0,9% pada suhu 31-34ºC. Selanjutnya oosit diaspirasi menggunakan syringe 3 ml dan jarum 23 G yang berisi Dulbeco's-Phosphate Buffer Saline (DPBS), kemudian dimaturasikan pada inkubator CO 2 modifikasi dalam medium Tissue Culture (CO 2 5%, kelembaban 99% dan suhu 37-39ºC). Oosit dibagi menjadi 2 kelompok yaitu kelompok kontrol (TCM-199) dan kelompok perlakuan (TCM-199 + 10% FCS). Angka maturasi dianalisa dengan Chi-Square, sedangkan kualitas oosit dianalisis secara deskriptif. Maturasi oosit sapi pada kelompok perlakuan berbeda nyata (P≤0,05) dibandingkan kelompok kontrol (55,22% vs 40,09%). Penggunaan 10% FCS pada medium maturasi dapat menghasilkan kualitas oosit matur yang lebih baik dibandingkan kelompok kontrol. Kesimpulan penelitian ini adalah penggunaan 10% FCS suplementasi dapat meningkatkan kemampuan maturasi oosit sapi in vitro. (Kata kunci: Fetal calf serum, Kultur oosit, Maturasi in vitro, Oosit sapi) ABSTRACT This study was conducted to investigate the utilization of ovaries from the slaughterhouse as oocytes source for in vitro fertilization (IVF). Fetal calf serum (FCS) was used as supplement of in-vitro maturation medium in order toincrease the successfullness of IVF. Ovaries were collected from local slaughterhouse and transported to the laboratory using 0.9% NaCl medium at 31-34ºC. The oocytes were aspirated by using a 3 ml syringe and 23 G needle containing Dulbeco's-Phosphate Buffer Saline (DPBS), then were maturated in modified CO 2 incubator (99% humidity and a temperature of 37-39ºC) in Tissue Culture . Oocytes were divided into 2 groups: control group

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Developmental potential of bovine oocytes cultured in different maturation and culture conditions

Cited by 73 publications

References 40 publications

Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos

Acetylation Level of Histone H3 in Early Embryonic Stages Affects Subsequent Development of Miniature Pig Somatic Cell Nuclear Transfer Embryos

The in vitro growth and maturation of follicles

Pengaruh Suplementasi Fetal Calf Serum Terhadap Kemampuan Maturasi in Vitro Oosit Sapi

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