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. 2006 Mar;23(3):121-7.
doi: 10.1007/s10815-006-9024-6. Epub 2006 May 13.

Sterile filtered paraffin oil supports in vitro developmental competence in bovine embryos comparable to co-culture

Jin Cheol Tae  1 , Eun Young Kim, Won Don Lee, Se Pill Park, Jin Ho Lim
Affiliations

Sterile filtered paraffin oil supports in vitro developmental competence in bovine embryos comparable to co-culture

Jin Cheol Tae et al. J Assist Reprod Genet. 2006 Mar.

Abstract

Purpose: To investigate whether sterile filtered light paraffin oil (SPO) overlaying is superior to washed light mineral oil (WMO) in supporting the in vitro developmental competence of bovine follicular oocytes. In addition, the effects of the two types of oil overlaying were compared with oil overlaying plus co-culture (CC) on bovine embryo development in vitro.

Methods: Bovine follicular oocytes retrieved from abattoir-derived ovary were in vitro matured, fertilized and cultured in 50 microL drops overlayed with WMO or SPO and were subsequently evaluated for development rates. In second experiment, day 2 embryos grown under WMO overlaying were further cultured for 6 days in the presence (WMO+CC and SPO+CC) or absence of adult ear skin fibroblast-based co-culture system overlaid with WMO or SPO. Blastocysts from each group were evaluated for total nuclei number or were further cultured for 48 h to evaluate post-hatching development.

Results: SPO overlaying resulted in significant higher (p < 0.05) development rate to morula (44.8% versus 30.6%) and blastocyst (32.8% versus 21.7%) than WMO. Also, treatment of the day 2 embryo cultures with SPO overlaying or oil plus CC (WMO+CC or SPO+CC groups) reached significantly higher development rates from the morula stage compared to embryo cultures treated with the WMO overlaying (p < 0.05). However, the development rates of the SPO treatment group (morula: 72.7%; blastocyst: 53.1%) were slightly high compared to development of the culture treated with WMO+CC (69.6 and 50.4%, respectively). This similar developmental competence pattern was also observed in cell number and embryo hatching rate.

Conclusion: SPO overlaying is superior to WMO and WMO+CC in supporting in vitro development of bovine embryos. The development rates are further enhanced when embryos are cultured in co-culture system overlaid with SPO. Thus, these data suggest that overlaying oil can significantly influence the pre-implantation embryo development in vitro.

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Figures

Fig. 1.
Fig. 1.
In vitro development of bovine day 2 embryos covered with washed light mineral oil (A, C, E and G) or sterile filtered light paraffin oil (B, D, F and H). Four- to eight-cell embryos (A and B) developed similarly until day 4 (8–16 cells; C and D) and then showed gradual differences in developmental capacity from day 6 (morula-blastocyst, E and F) to day 8 (G and H) according to oil variation. Sterile filtered paraffin oil supports increased and improved embryo production in vitro. Scale bars: 200 μm.
Fig. 2.
Fig. 2.
Differential stained inner cell mass and trophectoderm cells of bovine day 8 blastocysts developed from washed light mineral oil overlay (A), WMO overlay plus co-culture (B), sterile filtered light paraffin oil overlay (C) or SPO overlay plus co-culture (D). All embryos were healthy with numerous cell numbers. Scale bars: 100 μm.
See this image and copyright information in PMC

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