Immortalization and characterization of dermic fibroblasts by overexpression of hTERT

• Grant number: 24/20827-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2025
• End date: April 30, 2026
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Ernesto da Silveira Goulart Guimarães
• Grantee: Heloisa Genovez Alcoforado Carneiro
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Organ and/or tissue failure is responsible for 45% of annual deaths in the world. Tissue engineering is an attractive solution, because it allows the development of functional tissues for partial or total transplants or implants. Artificial tissues require functional cells, and primary cells are the gold standard for functionality, but they are not suitable for long in vitro cultivation. Transdifferentiation of dermal fibroblasts can be used to generate cells from different tissues, but their limited proliferative capacity, due to replicative senescence, hinders the genetic modification and long-term culturing required in this approach. The immortalization of fibroblasts before transdifferentiation can facilitate transfections and allow the generation of sufficient numbers of cells for application in tissue engineering. The aim of this study is to immortalize dermal fibroblasts by overexpressing hTERT and to characterize the lineages generated in terms of escape from senescence, cell cycle regulation/proliferation, and chromosomal alterations. The insertion of hTERT will be confirmed by qualitative PCR. The immortalized lineages will be characterized by flow cytometry, RT-qPCR and MLPA.