A novel technique for in vitro maturation of sheep oocytes in a liquid marble microbioreactor

PurposeThe aim of this work was to develop a microbioreactor using liquid marble (LM) as a novel system for oocyte in vitro maturation (IVM) in small volumes.MethodsCumulus-oocyte complexes (COCs) obtained from slaughterhouse sheep ovaries were in vitro matured in a LM system prepared by placing a drop (30 μl containing 10 COCs) suspended in TCM 199 supplemented with 10 % (v/v) oestrus sheep serum (OSS) and 0.1 IU FSH and LH onto a polytetrafluoroethylene (PTFE) particle bed (LM group). As a control group (CTRL group), COCs were in vitro matured in standard volume and conditions (600 μl of IVM medium in a four-well dish). After 24-h culture at 38.5 °C in 5 % CO2 in air, COCs were released from LM and the following parameters were evaluated: (a) percentage of MII oocytes, (b) oocyte developmental competence following in vitro fertilization (IVF) or parthenogenetic activation (PA) and embryo culture for 8 days in synthetic oviductal fluid (SOF) medium at 38.5 °C in 5 % O2, 5 % CO2, and 90 % N2.ResultsThe results indicated similar percentage of MII oocytes in LM and CTRL groups (88.0 vs. 92.0 %). No differences were observed in blastocyst rate after IVF (LM 47.5 % vs. CTRL 50.2 %, P=0.637) or PA (LM 44.4 % vs. CTRL 48.3 %, P=0.426).ConclusionsThe results indicate that LM microbioreactor is a viable technique that provides a suitable microenvironment to induce oocyte in vitro maturation.

[1]  Peggy P Y Chan,et al.  Cardiogenesis of Embryonic Stem Cells with Liquid Marble Micro‐Bioreactor , 2015, Advanced healthcare materials.

[2]  M. Pourghasem,et al.  Embryological Aspects of Oocyte In Vitro Maturation , 2013, International journal of molecular and cellular medicine.

[3]  H. R. Tervit,et al.  Successful culture in vitro of sheep and cattle ova. , 1972, Journal of reproduction and fertility.

[4]  C. Galli,et al.  Effect of prematuration, meiosis activating sterol and enriched maturation medium on the nuclear maturation and competence to development of calf oocytes. , 2004, Theriogenology.

[5]  Ying-pu Sun,et al.  Effect of culture medium volume and embryo density on early mouse embryonic development: Tracking the development of the individual embryo , 2012, Journal of Assisted Reproduction and Genetics.

[6]  Barry Behr,et al.  Metabolomic assessment of oocyte viability. , 2009, Reproductive biomedicine online.

[7]  Wei Shen,et al.  Respirable liquid marble for the cultivation of microorganisms. , 2013, Colloids and surfaces. B, Biointerfaces.

[8]  John J. Robinson,et al.  Conceptus growth and development following in vitro culture of ovine embryos in media supplemented with bovine sera , 1998 .

[9]  Sixue Chen,et al.  Treatment with the Proteasome Inhibitor MG132 during the End of Oocyte Maturation Improves Oocyte Competence for Development after Fertilization in Cattle , 2012, PloS one.

[10]  G. Leoni,et al.  Influence of vasoactive intestinal peptide (VIP), atrial natriuretic peptide (ANP) and insulin-like growth factor-I (IGF-I) on in vitro maturation of prepubertal and adult sheep oocytes , 1996, Zygote.

[11]  D. Gardner,et al.  Effect of incubation volume and embryo density on the development and viability of mouse embryos in vitro. , 1992, Human reproduction.

[12]  K. Chiba,et al.  Damage of embryo development caused by peroxidized mineral oil and its association with albumin in culture. , 2009, Fertility and Sterility.

[13]  K. Tanemura,et al.  Hanging drop monoculture for selection of optimal antioxidants during in vitro maturation of porcine oocytes. , 2014, Reproduction in domestic animals = Zuchthygiene.

[14]  K. Seow,et al.  Effects of growth factors and granulosa cell co-culture on in-vitro maturation of oocytes. , 2009, Reproductive biomedicine online.

[15]  N. Kawano,et al.  Delay of nuclear maturation and reduction in developmental competence of pig oocytes after mineral oil overlay of in vitro maturation media. , 2002, Reproduction.

[16]  David Quéré,et al.  Liquid marbles , 2001, Nature.

[17]  D. Izquierdo,et al.  Current status of in vitro embryo production in sheep and goats. , 2014, Reproduction in domestic animals = Zuchthygiene.

[18]  Sajal Gupta,et al.  Could oxidative stress influence the in-vitro maturation of oocytes? , 2009, Reproductive biomedicine online.

[19]  J. L. Hill,et al.  Development of ovine embryos in synthetic oviductal fluid containing amino acids at oviductal fluid concentrations. , 1996, Biology of reproduction.

[20]  N. Songsasen,et al.  Requirement for, and patterns of, pyruvate and glutamine metabolism in the domestic dog oocyte in vitro , 2007, Molecular reproduction and development.

[21]  S. Succu,et al.  Expression pattern of zygote arrest 1 (ZAR1), maternal antigen that embryo requires (MATER), growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) genes in ovine oocytes and in vitro-produced preimplantation embryos. , 2008, Reproduction, fertility, and development.

[22]  M. Gutiérrez,et al.  Mammalian cell cryopreservation by using liquid marbles. , 2015, ACS applied materials & interfaces.

[23]  J. Reinsberg,et al.  Pitfalls in assessment of progesterone production by granulosa cells cultured in contact with silicone rubber or paraffin oil , 2004, Archives of Gynecology and Obstetrics.

[24]  D. Gardner,et al.  Amino acids and ammonium regulate mouse embryo development in culture. , 1993, Biology of reproduction.

[25]  P. Loi,et al.  Development of parthenogenetic and cloned ovine embryos: effect of activation protocols. , 1998, Biology of reproduction.

[26]  Robert B Gilchrist,et al.  Oocyte maturation: emerging concepts and technologies to improve developmental potential in vitro. , 2007, Theriogenology.

[27]  M. Gupta,et al.  Epidermal growth factor can be used in lieu of follicle-stimulating hormone for nuclear maturation of porcine oocytes in vitro. , 2010, Theriogenology.

[28]  Yutao Du,et al.  The Well-of-the-Well system: an efficient approach to improve embryo development. , 2008, Reproductive biomedicine online.

[29]  G. Vajta,et al.  New method for culture of zona‐included or zona‐free embryos: The Well of the Well (WOW) system , 2000, Molecular reproduction and development.

[30]  S. M. Downs,et al.  Metabolism of radiolabeled glucose by mouse oocytes and oocyte-cumulus cell complexes. , 1999, Biology of reproduction.

[31]  W. Shen,et al.  Tumor Inside a Pearl Drop , 2012, Advanced healthcare materials.

[32]  Junfei Tian,et al.  Microreactors: Liquid Marbles as Micro‐bioreactors for Rapid Blood Typing (Adv. Healthcare Mater. 1/2012) , 2012 .

[33]  K. Koyama,et al.  In vitro maturation system for individual culture of bovine oocytes using micro-volume multi-well plate. , 2013, The Japanese journal of veterinary research.