Clonal Amniotic Fluid-Derived Stem Cells Express Characteristics of Both Mesenchymal and Neural Stem Cells1

Abstract

Recent evidence has shown that amniotic fluid may be a novel source of fetal stem cells for therapeutic transplantation. We previously developed a two-stage culture protocol to isolate a population of amniotic fluid-derived mesenchymal stem cells (AFMSCs) from second-trimester amniocentesis. AFMSCs maintain the capacity to differentiate into multiple mesenchymal lineages and neuron-like cells. It is unclear whether amniotic fluid contains heterogeneous populations of stem cells or a subpopulation of primitive stem cells that are similar to marrow stromal cells showing the behavior of neural progenitors. In this study, we showed a subpopulation of amniotic fluid-derived stem cells (AF-SCs) at the single-cell level by limiting dilution. We found that NANOG- and POU5F1 (also known as OCT4)-expressing cells still existed in the expanded single cell-derived AF-SCs. Aside from the common mesenchymal characteristics, these clonal AF-SCs also exhibit multiple phenotypes of neural-derived cells such as NES, TUBB3, NEFH, NEUNA60, GALC, and GFAP expressions both before and after neural induction. Most importantly, HPLC analysis showed the evidence of dopamine release in the extract of dopaminergic-induced clonal AF-SCs. The results of this study suggest that besides being an easily accessible and expandable source of fetal stem cells, amniotic fluid will provide a promising source of neural progenitor cells that may be used in future cellular therapies for neurodegenerative diseases and nervous system injuries.

Isolation, characterization, and in vitro differentiation of ovine amniotic stem cells

“…In recent years, several studies have demonstrated the ability to isolate amniotic membrane stem cells (AMSCs) and amniotic fluid stem cells (AFSCs) (In 't Anker et al 2003;Tsai et al 2004;Tsai et al 2006). These cells have phenotypes attributable to pluripotent cells and can differentiate into different cell types due to the three different embryonic layers (De Coppi et al 2007).…”

Preclinical regulatory validation of a 3-stage amniotic mesenchymal stem cell manufacturing protocol

“…It remains to be seen whether these alternatives would lead to any advantage, or disadvantage, in clinical-grade processing. Also, it remains to be determined whether amniotic stem cells recently obtained by somewhat different methods and described as more primitive than purely mesenchymal are actually the same cells that we and others refer to as AMSCs, which have been shown to be able to give rise to cells from more than one germ layer [13,14].…”

Isolation of Human Neural Stem Cells from the Amniotic Fluid with Diagnosed Neural Tube Defects

“…Our AFNSCs maintained or increased their expression of these markers during long-term cultivation in vitro. AF has been identified as an ideal source to isolate fetal multipotent stem cells (AFSCs) [24,39]. AFSCs also express some markers with mesenchymal stem cells, such as CD73, CD105, and CD117, and some pluripotent stem cell markers, such as Oct-4, Nanog, and Sox2.…”