Health & Environmental Research Online (HERO)


Print Feedback Export to File
3201191 
Journal Article 
A novel technique for in vitro maturation of sheep oocytes in a liquid marble microbioreactor 
Ledda, S; Idda, A; Kelly, J; Ariu, F; Bogliolo, L; Bebbere, D 
2016 
Yes 
Journal of Assisted Reproduction and Genetics
ISSN: 1058-0468
EISSN: 1573-7330 
33 
4 
513-518 
English 
26852233 
WOS:000373573200013 
PURPOSE: The 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.

METHODS: Cumulus-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.

RESULTS: The 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).

CONCLUSIONS: The results indicate that LM microbioreactor is a viable technique that provides a suitable microenvironment to induce oocyte in vitro maturation. 
Liquid marble; Bioreactor; Oocyte; In vitro maturation; Embryo development