Regenerative Medicine 2

Research title:

Placental progenitor cells and their derived acellular products for regenerative treatments to reverse ovarian failure in large animal models

Tutor: Prof. Fausto Cremonesi

Co-Tutor: Dott.ssa Anna Lange Consiglio

 

Contact details

Prof. Fausto Cremonesi, DVM

Department of Veterinary Medicine and Centro Clinico-Veterinario e Zootecnico-Sperimentale di Ateneo

Reproduction Unit

Via dell’Università, 6

26900 Lodi - Italy

This email address is being protected from spambots. You need JavaScript enabled to view it.

Tel: +39 02 503.31150/51/52/53

Fax: +39 02 503.31115

 

State of the art

The development of economic guidelines for increasing milk production has caused some negative effects, such as increase in metabolic disease at the transition period, lameness, and decrease in reproductive performance. Recent studies showed that on total culled cows, the frequency of reproductive disorders is mainly due to ovarian abnormalities. Of these abnormalities, the percentages of ovarian failure, cysts, and adhesions are 11.45%, 5.22%, and 6.38%, respectively. Such reproductive failures cause considerable economic damage in the dairy management.

Recent researches show that stem cell therapy has been used as an important research tool in human disease and regenerative medicine treatment and that mesenchymal stem cells (MSCs) can restore partial ovarian function after transplantation in mice with premature ovarian failure caused by chemotherapy. Our research group recently retrieved MSCs from extra-fetal tissues in large animal species comparing the proliferative and differentiative potential of amniotic membrane-derived MSCs (AMCs) with bone marrow derived MSCs together with their possible application in the treatment of spontaneous horse tendon injuries and endometritis. We also demonstrated the potential immunomodulatory properties of AMCs and their conditioned medium (AMC-CM) in vitro as well as proved the efficacy of AMC-CM in the treatment of spontaneous horse tendon injuries in vivo. Taken together, the AMCs features described so far by our group suggest this cell type as the most suitable for cellular transplantation approaches and its derived conditioned media (composed of soluble factor and microvesicles) as a novel, cell-free therapeutic product in regenerative medicine. Platelet rich plasma (PRP), composed of growth factors released by platelet, could represent another regenerative therapeutic approach for reproductive diseases as demonstrated by our research group in vitro and in vivo.

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Calf delivery at the barn and stripping of amnions from allantois in the lab.

 

Aim of the project

The present study is aimed to evaluate the efficacy of amniotic progenitor cells, their conditioned media and microvesicles in the therapy of cows with ovarian failure. Moreover, the effect of platelet rich plasma will also be investigated.

The project will be organized in different steps

1) Collection of bovine amniotic membrane, isolation and culture of amniotic-derived cells. Collection of CM and the isolation of microvesicles will also be carried out. Characterization and differentiation studies of amniotic cells, characterization of CM and evaluation of microvesicles, including the library of their micro RNA, will be main aims of this step. At the same time, bovine PRP will be produced by cows affected by ovarian failure and standardized for concentration of platelet and growth factors.

2) Identification of animals affected by ovarian failure based on clinical detection of anoestrus, ultrasound and hormonal assessments. These animals were randomly divided into control group and groups treated by AMCs, CM, microvesicles and PRP by ultrasound intraovarian injection. Ultrasound to identify change in the ovaries, and blood collection to detect serum estradiol, progesterone and Antimullerian hormone concentrations will be performed once weekly for six weeks before the treatment and eight weeks after the treatment.

3) Breeding experiments. Frozen semen of the same bull will be used to artificial inseminate animal eventually detected in oestrus. Pregnancy rate and live calves will be registered.

This project could represent the development of new strategies to counteract ovarian hypotrophy and other causes of hypofertility related to ovarian dysfunction in cows as well as represents a model for restoring fertility in women undergoing chemotherapy or anticancer treatments. 

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Recent publications of the tutor in the field

1) Perrini C., Strillacci MG.,Bagnato A., Esposti P., Marini MG., Corradetti B., Bizzaro D., Idda A., Ledda S., Capra E., Pizzi F., Lange-Consiglio A., Cremonesi F. Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in endometrial cells in an in vitro model Stem Cell Research and Therapy 2016, DOI: 10.1186/s13287-016-0429-6

2) Lange-Consiglio A., Cazzaniga N., Garlappi R., Spelta C., Pollera C., Perrini C., Cremonesi F. Platelet concentrate in bovine reproduction: effects on in vitro embryo production and after intrauterine administration in repeat breeder cows. Reproduction Biology and Endocrinology 2015 13:65.

3) Corradetti B., Correani A., Romaldini A., Marini M.G, Bizzaro D., Perrini C., Cremonesi F., Lange-Consiglio A. Amniotic membrane-derived mesenchymal cells and their conditioned media: potential candidates for uterine regenerative therapy in the horse. PLoS ONE, 2014, 9 (10) e111324.

4) Lange-Consiglio A., Rossi D., Tassan S., Perego R., Cremonesi F., Parolini O. Conditioned medium from horse amniotic membrane-derived multipotent progenitor cells: Immunomodulatory activity in vitro and first clinical application in tendon and ligament injuries in vivo. Stem Cells and Development. 2013; 22(22):3015-24.

5) Lange-Consiglio A., Tassan S., Corradetti B., Meucci A., Perego R., Bizzaro D., Cremonesi F. Investigating the efficacy of amnion-derived compared with bone marrow-derived mesenchymal stromal cells in equine tendon and ligament injuries. Cytotherapy. 2013; 15(8):1011-1020.