Biology Reference
In-Depth Information
Chapter 1
Nucleofection of Ovine Amniotic Fluid-Derived
Mesenchymal Stem Cells
V. Curini, A. Colosimo, A. Mauro, M. Turriani, A. Gloria, M. Mattioli,
and B. Barboni
Abstract Amniotic fluid has attracted increasing attention in recent years as a
possible source of stem cells. Amniotic stem cells have high differentiation ability
and low immunogenicity, and are thus an ideal candidate for stem cell-based
regenerative therapy. To assess their potential applicability, preclinical studies
have been initiated. In this context, the availability of GFP-expressing cells could
be extremely useful as a protein marker to visualize transferred stem cells within
damaged tissue. In the present study, nucleofection, a recent electroporation-based
technique, was used to transfect GFP-expressing plasmids into ovine amniotic
fluid-derived stem cells. The study shows that this transfection method can be
used to generate stable transgene expression in amniotic stem cells without altering
their differentiation potential.
Keywords Amniotic cells • Nucleofection • Ovine • Staminality markers
1.1
Introduction
In the field of regenerative medicine, stem cells represent a useful tool for restoring
the function of damaged tissues and organs. Recently, scientists have given special
attention to amniotic fluid-derived cells because these cells have a high degree of
plasticity, as they are able to differentiate in vitro into neurons, osteoblasts,
fibroblasts, adipocytes, chondrocytes, hepatocytes, and endothelial cells (Parolini
et al.
2009
). Immunocytochemistry and gene expression analyses confirmed the
presence of specific markers, such as OCT-4A, SOX2, NANOG, and TERT, as
V. Curini (
*
) • A. Colosimo • A. Mauro • M. Turriani • A. Gloria • M. Mattioli • B. Barboni
Department of Biomedical Comparative Sciences, University of Teramo, Teramo, Italy
e-mail:
valentinacurini@yahoo.it
