Effect of hyaluronic acid in a serum free maturation medium on bovine embryo development

Guienne, B. Marquant-Le; Guyader-Joly, C.; Ponchon, S.; Delalleau, Nathalie; Humblot, P.

doi:10.1016/s0093-691x(99)91945-3

Cited by 4 publications

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“…Durante a maturação, ocorre secreção de grandes quantidades de ácido hialurônico (HA) e outras proteínas que originam uma matriz extracelular gelatinosa envolvida na expansão do complexo cumulus oophorus-oócito [13,29]. Quando o HA é adicionado ao meio de maturação e cultivo há um aumento do potencial de desenvolvimento de oócitos [19] e embriões [8,32] bovinos. A expressão da proteína ácido hialurônico sintase 2 (HAS2), envolvida com a síntese de HA pelas células do cumulus, tem sido relacionada ao desenvolvimento da competência do oócito [7].…”

Section: Resultsunclassified

Expressão gênica das células do cumulus oophorus de bovinos após a vitrificação

Arend

Vieira

Cesaro

et al. 2018

Acta Scientiae. Vet.

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Um dos desafios da criobiologia continua sendo o desenvolvimento de um método que proporcione a manutenção da viabilidade após a criopreservação de oócitos imaturos na espécie bovina. A vitrificação tem sido a metodologia que proporciona resultados de sobrevivência após a criopreservação mais promissores para complexos cumulus-oócito (CCOs) imaturos bovinos. Entretanto, a ação dos crioprotetores sobre as células do cumulus oophorus, no que diz respeito à regulação da expressão de genes importantes nesta fase, ainda é pouco compreendida. O objetivo do trabalho foi avaliar a expressão gênica das proteínas ácido hialurônico sintase 2 (HAS2), link protein (HAPLN1), conexina 43 (GJA1) e proteína de choque térmico HSP70-1 (HSP70-1) em células do cumulus oophorus de oócitos imaturos bovinos submetidos a exposição e/ou vitrificação na solução crioprotetora (SV) composta por 20% de etileno glicol (EG) + 20% dimetil sulfóxido (DMSO) + 0,5M de sacarose. Os CCOs foram selecionados e distribuídos em 4 grupos experimentais: G1, CCOs não submetidos a maturação in vitro (MIV); G2, CCOs submetidos à MIV; G3, CCOs maturados após a exposição à SV; e G4, CCOs maturados após a vitrificação com a SV. A MIV foi realizada em TCM 199, suplementado com soro de égua em estro, à 39 o C, 5% de CO 2 e máxima umidade relativa, por 22 a 24 horas. A extração do RNA das amostras de células do cumulus foi realizada pelo método do fenol-clorofórmio seguido por uma etapa de captação específica do mRNA. Após a utilização da técnica de RT-PCR para a obtenção do cDNA e amplificação dos quatro fragmentos específicos, a análise dos resultados não mostrou diferença significativa entre os grupos para a abundância relativa dos transcritos de link protein (p=0,486), HAS2 (p=0,394), conexina 43 (p=0,116) e HSP70-1 (p=0,248).

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Section: Resultsunclassified

Expressão gênica das células do cumulus oophorus de bovinos após a vitrificação

Arend

Vieira

Cesaro

et al. 2018

Acta Scientiae. Vet.

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“…This may be partly due to follicular-synthesized components such as hyaluronan or midkine. Granulosa and expanding cumulus cells in the follicle secrete large amounts of hyaluronan, and when hyaluronan is added to the maturation and culture media, it improves the developmental potential of bovine oocytes and embryos (Marquant-Le Guienne et al, 2002) and the blastocysts' cell numbers (Stojkovic et al, 1999). Stojkovic et al (2002) found that blastocysts cultured in the presence of hyaluronan contained a similar total number of cells as in vivo derived embryos.…”

Section: Discussionmentioning

confidence: 99%

Oocyte maturation in the presence of randomly pooled follicular ﬂuid increases bovine blastocyst yield <i>in vitro</i>

Mohammed¹,

Karasiewicz²,

Papis³

et al. 2005

J. Anim. Feed Sci.

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The objective of the study was to examine preimplantation development of the zygotes derived from bovine oocytes matured in the presence of follicular fluid (bFF) randomly pooled from large (>10 mm in diameter) highly vascularised ovarian follicles originating from slaughter-house ovaries. The maturation medium was supplemented with 20% bFF. The controls comprised 20% bFF+10% FCS, and 10% FCS.The use of three separate batches of bFF have shown that the rates of cleavage (78-87%) and blastocyst formation (about 40%) obtained after bFF-supplemented maturation are similar or higher than those in the FCS control. In another experiment morphological selection of Grade I oocytes was performed after maturation or insemination. Selection of Grade I matured oocytes increased cleavage rate (to 85-91%) and blastocyst yield (to 43%). When Grade I inseminated oocytes were selected, then 92-93% of zygotes divided, and 47% reached blastocyst stage. Blastocyst rates were higher after bFF-supplemented maturation than after FCS-supplemented one, both in Grade I matured (43.0 as opposed to 40.7%) and Grade I inseminated (47.3 against 45.8%) oocytes.The majority of all zygotes underwent the first cleavage division between 27 and 30 hpi. Zygotes originating from oocytes matured in the presence of bFF or bFF+FCS were found dividing until 48 hpi whereas those obtained from control oocytes no longer cleaved after 33 hpi.In conclusion, oocyte maturation in the presence of bovine bFF randomly pooled from large follicles of slaughter-house ovaries supports as high developmental capacity of presumed zygotes as does the FCS-control.KEY WORDS: bovine follicular fluid, in vitro maturation, in vitro insemination, cleavage, blastocyst rate

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“…In the present study, a high concentration of HA (molecular weight: 1.69 ϫ 10 6 ) was used to increase the viscosity of culture medium. Marquant-Le Guienne et al (1998) demonstrated that HA can successfully substitute for sera for in vitro maturation of bovine oocytes. In support of the results from the present study, Stojkovic et al (1999a) and several other studies have demonstrated that supplementation of culture medium with HA improves in vitro development of bovine (Furnus et al, 1998) and pig (Miyoshi et al, 1999) embryos *All embryos that were frozen in 1.5 mol ethylene glycol l -1 were recovered after thawing.…”

Section: Discussionmentioning

confidence: 99%

Effects of high concentrations of hyaluronan in culture medium on development and survival rates of fresh and frozen-thawed bovine embryos produced in vitro

Stojkovic¹

2002

Reproduction

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Hyaluronic acid (HA) is the main glycosaminoglycan present in follicular, oviductal and uterine fluids. The main functions of HA include dynamic processes that are mediated through interaction with extracellular matrix components, regulation of gene expression, cell proliferation and cell differentiation. HA increases the viscosity of solutions and also has several physiological functions, including regulation of water distribution and waterbinding capacity. The addition of 6 mg HA ml-1 to synthetic oviduct fluid (SOF; SOF-HA) culture medium on day 5 (IVF = day 0) significantly (P < 0.001) increased the viscosity of the medium in comparison with SOF culture medium containing BSA (SOF-BSA). On day 8, rate of blastocyst development in SOF-HA culture medium was significantly (P < 0.05) higher than in SOF-BSA culture medium (38.2 versus 29.3%). The number of trophectoderm cells and the total number of cells of expanded blastocysts cultured in the presence of HA were significantly (P < 0.01) higher in comparison with expanded blastocysts cultured in the presence of BSA (88.9 ± 7.3 versus 67.6 ± 3.0 and 130.1 ± 10.9 versus 104.8 ± 2.5, respectively). After freezing and thawing, the percentage of day 8 embryos that re-expanded and hatched when cultured with SOF-HA was greater than that of embryos cultured with SOF-BSA (11.3 and 10.5% versus 75.5 and 36.8%, respectively). After thawing, the ATP contents of in vivo-derived, SOF-HA and SOF-BSA expanded blastocysts were similar. The embryos cultured with HA showed less ultrastructural deviation and de-differentiation after freezing and thawing than the embryos cultured with BSA. This study demonstrates that HA improves the developmental capacity of bovine embryos under in vitro conditions and is warranted as a supplement for in vitro production of bovine embryos, particularly if they are to be cryopreserved.

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Effect of hyaluronic acid in a serum free maturation medium on bovine embryo development

Cited by 4 publications

References 0 publications

Expressão gênica das células do cumulus oophorus de bovinos após a vitrificação

Expressão gênica das células do cumulus oophorus de bovinos após a vitrificação

Oocyte maturation in the presence of randomly pooled follicular ﬂuid increases bovine blastocyst yield <i>in vitro</i>

Effects of high concentrations of hyaluronan in culture medium on development and survival rates of fresh and frozen-thawed bovine embryos produced in vitro

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