In recent decades, assisted reproductive techniques have been developed and have been mainly used in livestock such as cattle, horses, pigs and small ruminants.
The research and commercial activity carried out at LRBA includes in particular the in vivo collection of oocytes and embryos, oocytes in vitro maturation, in vitro fertilization (IVF) or assisted fertilization (intracytoplasmic sperm injection, ICSI), in vitro embryo culture. Furthermore, protocols for the cryopreservation of gametes and embryos (traditional freezing and vitrification in straw or devices such as cryoloop) have been developed at our facility.
Oocytes and embryos can also be used for the development of toxicological tests of drugs and chemicals.
SELECTED PAPERS
1. Merlo B, Del Prete C, Mari G, Iacono E. Overnight holding aids in selection of developmentally competent equine oocytes. Anim Reprod Sci. 2022 Oct;245:107071. doi: 10.1016/j.anireprosci.2022.107071. Epub 2022 Sep 17.
2.Del Prete, Chiara; Calabria, Alfonso; Longobardi, Valentina; Palumbo, Veronica; Merlo, Barbara; Iacono, Eleonora.; Tafuri, Simona; Carotenuto, Domenico; Ciani, Francesca; Damiano, Sara; Ciarcia, Robertoa; Cocchia, Natascia. Effect of Aqueous Extract of Maca Addition to an Extender for Chilled Canine Semen. Animals 2022; 12(13); Article number 1638.
3. Gonzalez-Plaza, Alejandroa; Brullo, Cristiano; Cambra, Josep M.; Garcia, Manuela; Iacono, Eleonora; Parrilla, Inmaculada; Gil, Maria Antonia; Martinez, Emilio A.; Martinez, Cristina A.; Cuello, Cristina. Equilibration time with cryoprotectants, but not melatonin supplementation during in vitro maturation, affects viability and metaphase plate morphology of vitrified porcine mature oocytes. Reprod Domest Anim 2022. Online ahead of print.
4. Dell’Eva G, Bolognini D, Iacono E, Merlo B. Superovulation protocols for dairy cows bred with SexedULTRA™ sex-sorted semen. Reprod Domest Anim 2019; 54(5):756-761.
5. Merlo B, Mari G, Iacono E. In vitro developmental competence of horse embryos derived from oocytes with a different corona radiata cumulus-oocyte morphology. Anim Reprod Sci 2018; 198: 233-237
6. Merlo B, Iacono E, Bucci D, Spinaci M, Galeati G, Mari G. Beta-mercaptoethanol supplementation of in vitro maturation medium does not influence nuclear and cytoplasmic maturation of equine oocytes. Reprod Dom Anim 2019; 51: 992-996.
7. Iacono E, Merlo B, Rizzato G, Mislei B, Govoni N, Tamanini C, Mari G. Effects of repeated transvaginal ultrasound-guided aspirations performed in anestrous and cyclic mares on P4 and E2 plasma levels and luteal function. Theriogenology 2014;82(2):225-231.
8. Zaniboni A, Merlo B, Zannoni A, Bernardini C, Lavitrano M, Forni M, Mari G, Bacci ML. Expression of fluorescent reporter protein in equine embryos produced through intracytoplasmic sperm injection mediated gene transfer (ICSI-MGT). Anim Reprod Sci 2013;137(1-2):53-61.
9. Merlo B, Iacono E, Regazzini M, Zambelli D. Cat blastocysts produced in vitro from oocytes vitrified using the cryoloop technique and cryopreserved electroejaculated semen. Theriogenology. 2008;70(1):126-130.
SELECTED ABSTRACTS
1. Gorlani C, Merlo B, Iacono E. Effects of myo-inositol on oocyte in-vitro maturation and subsequent embryo development in cows. Accepted for Annual Meeting of European College in Animal Reproduction.
2. Merlo B, Mari G, Iacono E. In vitro developmental competence of horse oocytes with different cumulus morphology. Proceedings of 72nd meeting of Italian Society of Veterinary Sciences 2018: 205.
3. Boccia L, Iacono E, Rossi B, Merlo B. Effetcs of hot season on bovine oocyte quality: how to byopass the poor oocytes quality during his season. Reprod Fertil Dev 2015; 27: 229.
4. Merlo B, Mislei B, Bulfone G, Rizzato G, Iacono E, Mari G. Vitrification of equine expanded blastocysts. Proceedings of 11th Congress of Italian Society of Animal Reproduction (S.I.R.A.), Giugno 2013: 102-105.
5. Merlo B, Iacono E, Mari G. In vitro survival and viability of bovine morulae after vitrification in cryoloop. Proceedings of the 2nd Cost Action FA0702, Gemini General Conference Maternal Interactions with gametes and Embryos, October 2009: 75 (Abstract).
Since 2006, research at LRBA have been focusing on the isolation, expansion, characterization and cryopreservation of mesenchymal stem cells (MSCs) from adult tissues (adipose tissue, bone marrow and peripheral blood) and fetal derived tissues (amniotic fluid, cord blood, Wharton's jelly and amniotic membrane), in different animal species (horse, cattle, pig, dog and cat). All projects aim to develop protocols that can be used for regenerative therapy.
The characterization of isolated cells implies the determination of the presence of stemness markers by flow cytometry, immunocytochemistry, real time PCR, and in vitro differentiation in mesodermal lines (bone, cartilage, adipose tissue). Ongoing studies are investigating gene expression of pro and anti-inflammatory cytokines. Moreover, morphological studies in order to verify the secretion of microvesicles usable for regenerative therapy were carried out. Studies on the metabolism of adult and fetal derived mesenchymal stem cells to assess their respiratory capacity and causes of early cellular aging are taking place.
PROJECTS FUNDED:
- POR FESR 2014-2020. Azione 1.2.2. Biomateriali multifunzionali per l’autoriparazione di tessuti e organi.
- PRIN 2009 Induzione e determinazione della pluripotenza di cellule embrionali, fetali e somatiche-mediante transfezione di geni pluripotenti (iPSCs)- nel bovino e nel cavallo.
- COFIN 2005 Isolamento e caratterizzazione di cellule staminali mesenchimali equine da midollo osseo e tessuto adiposo; potenziale impiego in un modello di lesione tendinea.
- PRRIITT 2004 Cellule staminali per riparazione tissutale: sviluppo di prodotti e protocolli per laboratori di ricerca.
SELECTED PAPERS
1. Lanci A, Merlo B, Mariella J, Castagnetti C, Iacono E. Heterologous Wharton’s Jelly Derived Mesenchymal Stem Cells Application on a Large Chronic Skin Wound in a 6-Month-Old Filly. Frontiers Vet Sci 2019;6: Art 9.
2. Iacono E, Pascucci L, Bazzucchi C, Cunto M, Ricci F, Rossi B, Merlo B. Could hypoxia influence basic biological properties and ultrastructural features of adult canine mesenchymal stem /stromal cells? Vet Res Comm 2019; 42: 297-308.
3. Merlo B, Teti G, Mazzotti E, Ingrà, L, Salvatore V, Buzzi M, Cerqueni G, Dicarlo M, Lanci A, Castagnetti C, Iacono E. Wharton’s Jelly Derived Mesenchymal Stem Cells: Comparing Human and Horse. Stem Cell Rev Rep 2019; 14: 574-584.
4. Iacono E, Pascucci L, Rossi B, Bazzucchi C, Lanci A, Ceccoli M, Merlo B. Ultrastructural characteristics and immune profile of equine MSCs from fetal adnexa. Reproduction 2019; 154: 509-519.
5. Iacono E, Lanci A, Merlo B, Ricci F, Pirrone A, Antonelli C, Mariella J, Castagnetti C. Effects of Amniotic Fluid Mesenchymal Stem Cells in carboxymethyl cellulose gel on spontaneous pressure sores healing: clinical outcome in seven hospitalized neonatal foals. Turk J Biol 2016; 40: 484-492. doi:10.3906/biy-1507-147.
6. Iacono E, Rossi B, Merlo B. Stem cells from foetal adnexa and fluid in domestic animals: an update on their features and clinical application. Reprod Domest Anim 2015;50:353-364. doi: 10.1111/rda.12499.
7. Iacono E, Merlo B, Romagnoli N, Rossi B, Ricci F, Spadari A. Equine Bone Marrow and Adipose Tissue Mesenchymal Stem Cells: Cytofluorimetric Characterization, In Vitro Differentiation, and Clinical Application. J Eq Vet Sci 2015; 35:130-140.
8. Rossi B, Merlo B, Colleoni S, Iacono E, Tazzari PL, Ricci F, Lazzari G, Galli C. Isolation and in vitro characterization of bovine amniotic fluid derived stem cells at different trimesters of pregnancy. Stem Cell Rev. 2014 Oct;10(5):712-24.
9. Iacono E, Merlo B, Pirrone A, Antonelli C, Brunori L, Romagnoli N, Castagnetti C. Effects of mesenchymal stem cells isolated from amniotic fluid and platelet-rich plasma gel on severe decubitus ulcers in a septic neonatal foal. Res Vet Sci 2012;93(3):1439-40.
10. Iacono E, Cunto M, Zambelli D, Ricci F, Tazzari PL, Merlo B. Could fetal fluid and membranes be an alternative source for mesenchymal stem cells (MSCs) in the feline species? A preliminary study. Vet Res Commun 2012;36(2):107-118.
11. Iacono E, Brunori L, Pirrone A, Pagliaro PP, Ricci F, Tazzari PL, Merlo B. Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse. Reproduction 2012;143(4):455-468.