Production of multipotent stromal cells (MSC) derived proteins for clinical application in veterinary medicine

• Grant number: 18/08295-3
• Support Opportunities: Research Grants - Innovative Research in Small Business - PIPE
• Start date: February 01, 2019
• End date: July 31, 2021
• Field of knowledge: Agronomical Sciences - Veterinary Medicine
• Agreement: FINEP - PIPE/PAPPE Grant
• Principal Investigator: Marina Landim e Alvarenga
• Grantee: Marina Landim e Alvarenga
• Company: Omics Biotecnologia Animal Ltda
• CNAE: Atividades veterinárias
• City: Botucatu
• Pesquisadores principais:( Últimos ): Fernanda da Cruz Landim
• Pesquisadores principais:( Antigos ): Danielle Jaqueta Barberini
• Associated scholarship(s): 20/00485-8 - Production of protein concentrate derived from multipotent stromal cells (MSC) for use in veterinary medicine, BP.TT

Abstract

Multipotent stromal cells (MSCs) are cells that have the ability to self-renew and differentiate in vitro in various cell lines, making them attractive for regenerative medicine, tissue engineering and cell therapy. One of the main mechanisms by which MSCs act on tissue regeneration is related to their paracrine activity, through the release of several cytokines, antioxidants and trophic and growth factors, creating a favorable microenvironment capable of modulating immune and inflammatory responses. In this sense, the evaluation of the protein content of the conditioned medium secreted by these canine cells seeks to understand its properties and possible applications in regenerative medicine. Several studies have demonstrated the effectiveness of the use of the secretoma produced by MSCs for the treatment of renal, bone, neurological, dermatological, hepatic, pulmonary and histopathological lesions. However, large-scale production in bioreactors and the composition of the secretoma under these culture conditions are not yet known, preventing the widespread use in regenerative therapy. Therefore, the objective of this project is to produce and analyze the secretoma of canine MSCs obtained from adipose tissue cultured in bottles and Spinner bottle-type bioreactors, with the objective of creating a product to be used in therapeutic protocols for the treatment of degenerative canine diseases. Four experiments will be performed. In the first we will compare the productivity and cellular viability in static culture and in spinner bottles using two types of microcarriers. In this study will be measured the total protein concentration, the production of lactic acid and ammonia by the cells. In a second experiment, the stability and bacterial contamination of the medium conditioned by the MSCs cultured in experiment 1 will be evaluated after cooling, freezing and lyophilization. The evaluation of the conditioned medium will be performed in experiment 3 by means of matched nanocromatography and mass spectrometry combined with isobaric labeling. In the experiment 4, a safety test will be carried out to evaluate the feasibility of the clinical application of the medium containing the MSM secretoma in the canine species. The results obtained will provide important information for a better understanding of the behavior of MSCs cultured in bottles and in bioreactors; as well as for the development of new strategies in regenerative therapies in which it would not be necessary to apply the cells, but rather the proteins produced by them, creating therapeutic possibilities more controlled and with reduced side effects. (AU)