Isolation and molecular characterization of amniotic fluid-derived mesenchymal stem cells obtained from Caesarean sections

Human amniotic fluid cells (AFCs) are immune-privileged with low immunogenicity and anti-inflammatory properties. Furthermore, they are high proliferative and have a broad differentiation potential, making them amenable for cell replacement therapies. Amniotic fluid (AF) is routinely obtained via amniocentesis. It contains heterogeneous populations of foetal-derived differentiated and undifferentiated progenitor cells including mesenchymal stem cells (MSCs). AF-derived mesenchymal stem cells (AF-MSCs) can self-renew and have a high proliferation and differentiation potential. In this study, we isolated AFCs from AF obtained during Caesarean sections (C-sections) and characterized them. The AFCs showed typical MSCs characteristics in relation to morphology, in vitro differentiation potential, cell surface marker expression and secreted factors. Subpopulations of AF-MSCs expressed several pluripotency-associated markers such as stage specific embryonic antigen 4 (SSEA4), c-Kit, TRA-1-60 and TRA-1-81, making them reprogrammable to induced pluripotent stem cells (iPSCs) without the use of ectopic gene expression. Additionally, they express the mesenchymal marker - Vimentin and multipotency-associated stem cell marker - CD133. Furthermore, the transcriptome and secretome analyses showed significant overlap with bone marrow-derived MSCs. C-section-derived AFMSCs can be routinely obtained without any risk to the foetus. Patient-specific AF-MSCs can be used for personalized cell therapies and disease modelling. The transcriptomes of human AF-MSCs from C-sections were compared to human fetal MSCs as well as human pluripotent cells (H1, H9, B4) by microarray analysis (PrimeView Human Gene Expression Array, Affymetrix, Thermo Fisher Scientific).