+ Translate

The use of sodium hyaluronate in freezing media for bovine and murine embryos

, : The use of sodium hyaluronate in freezing media for bovine and murine embryos. Cryobiology 30(2): 172-178

Four experiments were designed to determine whether sodium hyaluronate (SH) may be used to replace newborn calf serum (NSC) in murine and bovine embryo freezing media. A total of 780 mouse and 178 cattle embryos were frozen. Freezing media were prepared in Dulbecco's phosphate-buffered saline supplemented with 20% NCS and 10% glycerol (v/v) (medium A). In experiments I and II, NCS was replaced by 0.1% SH (w/v) in medium B and by 0.1% polyvinyl alcohol (PVA) (w/v) in medium C. In experiments III and IV, NCS was replaced in mouse embryo freezing medium by three different molecular weights of SH as follows: 0.1 or 0.2% (SH-1; lt 3 times 10-5 Da) in medium B, by 0.1 or 0.05% (SH-2; 5-7.5 times 10-5 Da) in medium C, and by 0.1 or 0.025% (SH-3; gt 1.2 times 10-6 Da) in medium D. Embryos were frozen and thawed using standard procedures. Glycerol was diluted from embryos after thawing in a single step with 1.0 M sucrose. After 48 h in culture (Ham's F-10 medium for cattle embryos and BWW medium for mouse embryos), embryos were evaluated for development to expanded or hatched blastocysts. Survival rates were compared by chi-2 analysis. In experiment I, there were no significant differences in mouse embryo survival or development between NCS, SH, and PVA groups (84.1, 79.2, and 83.3%, respectively). In experiment II, there were no significant differences in bovine embryo survival between NCS and SH groups (67.2 and 67.7%), but both had significantly higher survival than the PVA group (37.9%; P lt 0.05). In experiment III, there was a significantly lower (P lt 0.05) embryo survival rate in the low-molecular-weight SH-1 group (18.6%) than in medium (SH-2)- or high (SH-3)-molecular-weight groups (78.6 and 92.5%, respectively). Although doubling the concentration of SH-1 in experiment IV resulted in improved embryo survival (63.4%), survival rates were significantly higher (P lt 0.05) in the medium-molecular-weight (SH-2; 82.3%) and high-molecular-weight (SH-3; 86.0%) groups despite reduced concentrations of these compounds in the freezing medium. Results suggest that SH can be used to replace biological sera in embryo freezing media.

Accession: 002530604

PMID: 8319487

DOI: 10.1006/cryo.1993.1016

Download PDF Full Text: The use of sodium hyaluronate in freezing media for bovine and murine embryos

Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:

Related references

Hasler, J.F., 2010: Synthetic media for culture, freezing and vitrification of bovine embryos. Media designed for the recovery, holding and cryopreservation of bovine and equine embryos are available from several commercial sources. In years past, some of these media contained bovine serum, although inclusion of serum in embryo transfer med...

Nowshari, M.A.; Brem, G., 1999: Refreezing of bovine and murine embryos by rapid-freezing procedure. Theriogenology 51(1): 172, Jan 1

Rondeau, M.; Guay, P.; Goff, A.K.; Cooke, G.M., 1996: Growth and metabolism of murine and bovine embryos in bovine uterine flushing-supplemented culture media. The aim of this study was to compare the development and metabolic activity of cultured murine and bovine embryos in 2 standard media (HAM F-10 and RPMI) in the presence or absence of bovine uterine flushings. Murine morulae (n = 653) and day 7 bo...

Vasuthevan, S.; Ng, S.C.; Edirisinghe, R.; Bongso, A.; Ratnam, S., 1992: The evaluation of various culture media in combination with dimethylsulfoxide for ultrarapid freezing of murine embryos. Bicarbonate-buffered HTF medium, Medicult, and T6 are as effective as PB1 medium when used in combination with DMSO in ultrarapid freezing of two-cell mouse embryos. However, the use of phosphate-buffered T6 results in reduced in vitro development...

Ambrosi, V.; Morini, G.; Parmigiani, E.; Bigliardi, E., 2002: Slow freezing of micromanipulated bovine embryos: recent progress in media composition. This study briefly describes the most common techniques and media used for slow freezing of bovine embryos and the principal characteristics of new media for freezing micromanipulated embryos.

Bondioli K.R.; Mertes P.C., 1985: The effect of calcium levels in the freezing media on in vitro survival of bovine embryos. Theriogenology 23(1): 181

Menino, A.J.; Cheek, H.; Takahara, L., 1983: Development of murine embryos in culture media supplemented with bovine serum albumin, bovine uterine fluid and heat-treated bovine serum. Proceedings of the annual meeting American Society of Animal Science Western Section4(34): 252-255

A.L.Long; Xiao Mei; Feng XiuLiang; Dou ZhongYing; Qiu Huai; Yang Qi; Lei AnMin; Yang ChunRong; Gao ZhiMin, 2002: Research on growth behavior of embryos for bovine and murine on primary murine embryos fibroblast cell feeder layer. The difference in growth behavior between bovine embryos and murine embryos was studied on PMEF (primary murine embryos fibroblast) feeder layer. The results showed as follows: With embryos having attached, bovine embryonic trophoblast formed a tr...

Lai, A.C.H.; Ryan, J.P.; Sanders, D.M., 1993: Decrease in cell volume of mouse oocytes/pronuclear embryos in freezing media/survival rates following ultrarapid freezing. Survival rates of ultrarapidly frozen mouse oocytes are much reduced compared with that of pronuclear stage embryos (PN). A major difference between fertilized and unfertilized mouse ova is that fertilized ova are more permeable to dimethyl sulfox...

Holy L., 1988: Freezing of bovine embryos i effect of freezing and thawing on quality and developmental stages. Revista Cubana De Ciencias Veterinarias: 183-192