Generation and characterization of Mesenchymal Stem Cells for use is cell therapy

Cell therapy, therapy in which cells are used for treatment, is the subject of intense research and presents great potential. Specifically, mesenchymal stem cells (MSC) are emerging as an effective and safe cell-based therapy for the treatment and prevention of many infectious and inflammatory conditions affecting humans and animals. MSCs are non-hematopoietic multipotent stem cells, belonging to the mesoderm layer which is one of the three germ layers. MSCs are most frequently derived from adult tissue sources such as bone marrow and adipose tissue or from birth associated tissue such as placental tissue, amnion, umbilical cord and cord blood, and poses the ability of multi-lineage differentiation into cell types such as adipocytes, osteoblasts, chondrocytes, myocytes, β-pancreatic islets cells and neuronal cells.

In our lab we generate MSCs from bovine umbilical cord, placenta and adipose tissue and murine bone marrow, and characterize them by testing their immunomodulation, their ability to differentiate into adipocytes and osteoblasts, we test their gene expression profile by means of RT-PCR and RNA seq and their epigenetic landscape by bisulfite sequencing and Chromatin Immunoprecipitation (ChIP).

Following systemic and local administration, allogeneic MSCs were shown to target inflamed tissues and to improve cure by immunomodulation and better bacterial clearance. We propose here to study the safety, feasibility and efficacy of MSCs for the treatment of inflammatory diseases such as mastitis and metritis (udder and uterine inflammation) in dairy cows. We hypothesize that allogeneic MSCs will populate inflamed organs and will alleviate inflammation and improve bacteriological cure.

Mastitis, an inflammatory response of the mammary tissue to invading pathogenic bacteria, is a major animal health and welfare problem in the dairy industry and is responsible for multibillion dollar of economic losses. Although improved hygiene and management techniques reduced and even eradicated some forms of the disease, mastitis still prevail in all dairy farms. Currently, the administration of antibiotics is the most common method of treatment and prevention of mastitis. However, this strategy has many disadvantages including low cure rate, increasing occurrence of bacterial resistance, and the presence of antibiotics residues in the milk. Therefore, alternative effective approaches for management of bovine mastitis should be looked for and tested.

(A) Changes in bWJ-MSC marker expression during culture time and passages. Note the decrease at p5. CD45 – leucocytes marker. Ctrl – PSMB gene. (B) Co-culture of bovine MSC (BUC – bovine umbilical cord, WJ – Warton jelly, at different passages) with peripheral blood cells inhibit the activation and proliferation of T cells. (C) bUC-MCS (green) in a cross talk with mammary epithelial cells (red – actin filaments, blue – DNA). (D) bUC-MSC (error head) exchange mitochondria (red) and cytoplasmic material (green) with epithelial cells