Conditioned culture medium developed with human mesenchymal stem cells

• Grant number: 21/00585-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2021
• End date: July 31, 2022
• Field of knowledge: Engineering - Biomedical Engineering - Bioengineering
• Principal Investigator: Elenice Deffune
• Grantee: Giullia Cavicchioli Barbosa
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The cell culture directed to both pharmaceutical and therapeutic areas can be made in different culture media, such as the serum-free MesencultÔ and those with the addition of Fetal Bovine Serum. Emerging in this concept, the Conditioned Medium Culture obtained through mesenchymal stem cells (MSC) culture has been shown as a good alternative considering the transplantation of cells cultivated can benefit from tissue's local response to the molecules that are been secreted, thus avoiding complications from cellular grafting or transplantation. This medium is, therefore, produced by the MSCs' secretion, whose cell type can be obtained through different sources, such as Human Umbilical Cord, which is usually thrown away after birth; these cells are characterized for their capacity or self-renew, proliferation, and differentiation in many cell lineage (multipotentiality), which does have huge importance to the science world, looking forward to its regenerative effect and factors' secretion that is responsible for both intercellular communication and signalization. Analyzing the secreted factors, Extracellular Vesicles (VEs) are found, which are known as nanometric particles able of carrying cytoplasmatic components (such as micro RNAs, messenger RNAs, cytokines, among others), that will then be released into the intercellular medium, being able to regulate different physiologic processes, like cell proliferation, differentiation, and migration. The VEs have labored characterization, however, they can be differentiated according to their size, surface markers, and origin, which allocate them in three subcategories: exosomes, microvesicles, and apoptotic particles. The study about MSCs and VEs relation has as goal show the importance when considering the therapeutic potential and medicine tool, thus characterizing VEs secreted by MSCs, producing conditioned medium from this cellular lineage and the comparisons between VEs obtained through three different culture media (nourished with fetal bovine serum, MSCs' secreted factors and MesenCultÔ medium) will assist in understanding, classification and identifying of those VEs, as well as verify the paracrine factors efficiency of paracrine factors arising from MSCs within the scope of cell therapy, considering yet higher precision and compatibility to human cells. In this project, there will be used techniques for defrosting cryopreserved cells, applying the three different conditions mentioned and the characterization by VEs Electronic Microscopy of Transmission, determination of both VEs' size and concentration, by the use of NanoSightÔ equipment, which will perform the analysis and tracking of these nanoparticles.