Investigating Amniotic Fluid Stem Cells Epithelial to Mesenchymal Transition in Culture

Abstract

Amniotic fluid plays a vital role in supporting the growth and development of the fetus. Different research studies have found the AF to have significant potential in treating diseases through regenerative medicine and tissue engineering. AF has also been of detrimental importance in diagnosing chromosomal abnormalities and neural tube defects, for which it’s been used for the past 40 years. The composition of the fluid is highly heterogeneous and highlights the presence of various proteins, cell cells, and stem cells. The aim of this project is to understand the identity of the Amniotic Fluid-derived Mesenchymal Stem Cells and prove that these originated through an Epithelial-Mesenchymal transition (EMT) process, happening in cultures over different time points. Four amniotic-fluid samples were collected for this research. The physician collected the samples using an amnio-drainage procedure and the samples were cultured using standard protocols. Once the cultures are established, flow cytometry was used to provide accelerated analysis of individual cells. The results showed that the expression of mesenchymal markers such as CD73 and CD90 start to elevate upon culture on Day 7 and keep increasing during the time points. In opposition, the AF cells exhibit reduced epithelial markers expression, shown by decreased detection of EpCAM. E-Cad expression begins to recover and increase by Day 14. In conclusion, AF mesenchymal stem cells are not inherently present in the amniotic fluid but only exist in an epithelial state in vivo. AFSC produces more mesenchymal proteins as they are cultured, undergoing EMT. These findings have implications for using AF stem cells in regenerative medicine and would allow an improved understanding of their fetal origins.