Reproduction 2

Research Title:

ENRICHED CULTURE SYSTEMS AND OOCYTE CRYOPRESERVATION IN ANIMAL MODELS 

Tutor: Prof. Gaia Cecilia LUVONI DVM, PhD, Dipl. ECAR

 

Contact details

Department of Health, Animal Science and Food Safety
University of Milan
Via Celoria, 10 - 20133 Milan, ITALY
Ph. +39 02 503 18147 Fax +39 02 503 18148
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State of the art

In vitro development of cryopreserved female gametes is still suboptimal. Cryopreservation induces negative effects on the germinal and the somatic compartments and "ad hoc" cultural microenvironments for frozen-thawed gametes might improve their chances to resume meiosis and develop into an embryo. It has been shown that enriched culture systems, as three-dimensional (3D) scaffolds or co-culture with companion cells restore the fertility potential of low competence fresh oocytes in different species (Pangas et al., Tissue Eng, 2003; 9: 1013-21; Vigo et al., Tissue Eng, 2005; 11: 709-714; Morselli et al., Reprod Domest Anim, 2017; 52: 83-8). The 3D systems are able to promote the physiological cell-to-cell communication throughout the maintenance of cell re-arrangement in a short or long-term in vitro culture, whereas the secreted factors of co-cultured companion cells have a pivotal role in cytoplasmic and nuclear maturation (Gilchrist et al., Hum Reprod Update, 2008; 14: 159-77). The encapsulation of granulosa cells in 3D matrix, as a functional follicle-like structure, may allow the self-organization, proliferation and endocrine functions of the cells, promoting their oocyte nursing capacities (Kossowska-Tomaszczuk et al., Tissue Eng. Part A, 2010; 16: 2063-73). This enriched system might provide the cultural conditions for a more efficient post-thaw survival of oocytes in terms of functional integrity and developmental competence.

 

Aims of the project

The project will be focused on the development of enriched culture systems for cryopreserved oocytes in animal models.

Specific aspects to investigate:

a) survival and proliferation rate of granulosa cells encapsulated in 3D scaffolds;

b) hormonal secretion of encapsulated granulosa cells;

c) ability of 3D granulosa cell systems to support the developmental competence of frozen/thawed oocytes.

 

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Recent publications of the tutor in the field

1. Morselli M.G., Canziani S., Vigo D., Luvoni G.C. (2017). A three-dimensional alginate system for in vitro culture of cumulus-denuded feline oocytes. Reproduction in Domestic Animals, 52: 83-8.

2. Morselli M.G., Luvoni G.C., Comizzoli P. (2016). The nuclear and developmental competence of cumulus-oocyte complexes is enhanced by three-dimensional coculture with conspecific denuded oocytes during in vitro maturation in the domestic cat model. Reproduction in Domestic Animals, doi: 10.1111/rda.12850.

3. Alves A.E., Padilha-Nakaghi L.C., Pires-Butler E.A., Apparicio M., Silva N.A.M., Motheo T.F., Vicente W.R.R., Luvoni G.C. (2016). Viability and growth of feline preantral follicles in vitro cultured with insulin growth factor and epidermal growth factor supplemented medium. Reproduction in Domestic Animals, doi: 10.1111/rda.12839.

4. Apparício M., MostachioG.Q., MotheoT.F., AlvesA.E., PadilhaL., Pires-Butler E.A., Savi P.A.P.,UscateguiR.A.R., LuvoniG.C., VicenteW.R.R. (2015). Distribution of cortical granules and meiotic maturation of canine oocytes in bi-phasic systems. Reproduction Fertility and Development, 27: 1082-7.

5. Apparicio M., RuggeriE., LuvoniG.C. (2013).Vitrification of immature feline oocytes with a commercial kit for bovine embryo vitrification.Reproduction in Domestic Animals, 48: 240-4.