regenerative medicine

Regenerative medicine is defined as an interdisciplinary field that aims at developing biological substitutes to restore and improve tissue functions following severe traumas. Over the last twenty years, significant progresses have been made in this field thanks to the use of adult mesenchymal stem cells thanks to their capacity to induce angiogenesis thus promoting tissue regeneration. Nevertheless, there is a growing body of evidence supporting the hypothesis that the enhancement of tissue regeneration is due mainly to the paracrine action exerted by hASCs and therefore to the pro-angiogenic and anti-apoptotic factors released in the extracellular environment.In our laboratory, we are currently working with human mesenchymal stem cells (hMSCs) and their conditioned media to demonstrate their in vivo pro-angiogenic potential when associated with a three-dimensional collagen scaffold INTEGRA® Flowable Wound Matrix. Specifically, the device, comprise of hMSC-conditioned medium, is loaded in the backside of BALB-C nude athymic mice. After an appropriate time, the scaffolds are harvested and analyzed to determine the presence of newly formed vessels and cellular components.

Despite the great potentiality that hMSCs represent in the field of regenerative medicine, there is a growing body of evidence suggesting that aging may affect hMSC properties such as the proliferation rate, the differentiation, as well as the regenerative potential and growth factor secretion. Therefore, the project aimed also to investigate this aspect evaluating the telomerase activity, the expression of specific genes involved in the oxidative stress and the glycolytic pathway, and the epigenetic modifications.

Methods:

  • Cell isolation and maintenance
  • Scaffold preparation and in vivo injection
  • DNA and RNA extraction
  • Retrotranscription
  • DNA and RNA electrophoresis
  • Qualitative and Real Time PCR
  • Sample processing for optical and electronic microscopy observation
  • Cell proliferation with BrdU
  • Telomere length evaluation
  • ELISA
  • Human Cytokine Array

Department of biotechnology and Life Science – DBSV