Amniotic-Fluid Stem Cells: Growth Dynamics and Differentiation Potential after a CD-117-Based Selection Procedure

Abstract

Amniotic fluid (AF) has become an interesting source of fetal stem cells. However, AF contains heterogeneous and multiple, partially differentiated cell types. After isolation from the amniotic fluid, cells were characterized regarding their morphology and growth dynamics. They were sorted by magnetic associated cell sorting using the surface marker CD 117. In order to show stem cell characteristics such as pluripotency and to evaluate a possible therapeutic application of these cells, AF fluid-derived stem cells were differentiated along the adipogenic, osteogenic, and chondrogenic as well as the neuronal lineage under hypoxic conditions. Our findings reveal that magnetic associated cell sorting (MACS) does not markedly influence growth characteristics as demonstrated by the generation doubling time. There was, however, an effect regarding an altered adipogenic, osteogenic, and chondrogenic differentiation capacity in the selected cell fraction. In contrast, in the unselected cell population neuronal differentiation is enhanced.

Human amniotic fluid-derived c-kit+ and c-kit− stem cells: growth characteristics and some differentiation potential capacities comparison

“…However, the c-kit ? cells were reported to constitute only 1-5% of the total AF cells (De Coppi et al 2007;Arnhold et al 2011). We obtained a similar result of 3.30 ± 1.24% by flow cytometric sorting.…”

“…A high purity AF cell population can be sorted using c-kit as a target, and it is considered that a purer cell population is safer for clinical use (Phermthai et al 2010). However, c-kit-based selection procedure may induce a different mesenchymal differentiation capacity from the unselected cell populations (Arnhold et al 2011). Stefano noted that the composition of AF cells has a precise timetable during gestation.…”

CD117+amniotic fluid stem cells

“…40 AFS cell isolation is based on a two-step protocol consisting in the prior immunological selection (via flow cytometry or magnetic cell sorting) of CD117 positive cells from the AF (approximately 1% of total AF cells) and in the subsequent expansion of these cells in culture. 35,37,[41][42][43] Isolated AFS cells can be expanded in feeder layer-free, serum-rich conditions without evidence of spontaneous differentiation in vitro. Cells are cultured in basic medium containing 15% of fetal bovine serum and Chang supplement.…”

Amniotic Fluid Progenitor Cells and Their Use in Regenerative Medicine

“…Amniotic fluid stem cells have been shown to be easily cultured and expanded upon collection and isolation De , Da Sacco et al, 2010, Kunisaki et al, 2007. De and Arnhold et al (Arnhold et al 2011) proved that after c-kit selection these cells are still exhibiting optimal growth rate. Their potential for differentiation has been proved in many published works and cells can be retrieved from different species like humansDe , Da Sacco et al, 2010, pigs (Zheng et al, 2010), goat Zheng et al, 2011 ), mouse and buffalo (Yadav et al, 2010).…”