Analysis of the induction potential for multidifferentiation of the secretome from dental pulp stem cell spheroid

• Grant number: 19/27492-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2020
• End date: December 31, 2022
• Field of knowledge: Health Sciences - Dentistry
• Principal Investigator: Katiúcia Batista da Silva Paiva
• Grantee: Daniel Barrozo Ferreira
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Studies indicate increased multi-differentiation potential of dental pulp stem cells (DPSCs) in the spheroid (3D) model. This is possibly linked to the phenomenon of central hypoxia, which occurs in spheroids after a specific size, but is not clear if the central hypoxia is an important parameter that influences the differentiation potential of the secretome, and if a hypoxic microenvironment also generates some modulation on these events. Thus, this paper intends to make an initial approach to the hypoxia influence on the DPSC's spheroid secretome, taking into account spheroid size/central hypoxia/multi-differentiation potential when grown in osteogenic medium. For this, we will generate 3 different sizes spheroids: 100 (without central hypoxia), 500 and 1.000 ¼m diameter. After 14 days in osteogenic medium, we will collect their conditioned medium and add them to undifferentiated DPSCs monolayer (2D). We will also do the opposite experiment, where the conditioned medium of 2D cells will be incubated in the spheroids. After 7 days in conditioned medium, we will analyze the differentiation markers in these cells by real time PCR. (AU)