Potential Role of Culture Mediums for Successful Isolation and Neuronal Differentiation of Amniotic Fluid Stem Cells

Abstract

In recent years, the use of stem cells has generated increasing interest in regenerative medicine and cancer therapies. The most potent stem cells derive from the inner cell mass during embryonic development and their use yields serious ethical and methodological problems. Recently, a number of reports suggests that another suitable source of multipotent stem cells may be the amniotic fluid. Amniotic fluid mesenchymal stem cells (AFMSCs) are capable of extensive self-renewal, able to differentiate in specialized cells representative of all three germ layers, do not show ethical restriction, and display minimal risks of teratomas and a very low immunogenity. For all these reasons, amniotic fluid appears as a promising alternative source for stem cell therapy. Their recent discovery implies a lack of knowledge of their specific features as well as the existence of a protocol universally recognized as the most suitable for their isolation, growth and long-term conservation. In this study, we isolated stem cells from six amniotic fluids; these cells were cultured with three different culture mediums [Mesenchymal Stem Cell Medium (MSCGM), PC-I and RPMI-1640], characterized by cytofluorimetric analysis, and then either frozen or induced to neuronal differentiation. Even if the immunophenotype seemed not to be influenced by culture medium (all six samples cultured in the above-mentioned mediums expressed surface antigens commonly found on stem cells), cells showed different abilities to differentiate into neuron-like cells and to re-start the culture after short\long-term storage. Cells isolated and cultured in MSCGM showed the highest proliferation rate, and formed neuron-like cells when sub-plated with neuronal differentiation medium. Cells from PC-I, on the contrary, displayed an increased ability to restart culture after short\long term storage. Finally, cells from RPMI-1640, even if expressing stem cells markers, were not able to differentiate in neuron-like cells. Further studies are still needed in order to assess the effective role of culture medium for a successful isolation, growth, differentiation and storage of AFMSCs, but our data underline the importance of finding a universally accepted protocol for the use of these cells.In the last few years, basic and clinical research carried out on the use of stem cells represents a revolution (I) in regenerative medicine and cancer therapies (2). Stem cells have an extensive capacity

Characterization and profiling of immunomodulatory genes in resident mesenchymal stem cells reflect the Th1-Th17/Th2 imbalance of psoriasis

“…As postulated, the most part of the genes encoding for Th2 cytokines in PSO-MSCs (CCL1, CCL22, CXCL12, IL2, IL3, IL4, IL13B, IL 22, IL 27, TGF-b1) are not downor up-regulated, confirming previous results on differentiated skin cells: psoriasis is characterized by a cytokine production profile polarized toward Th1-Th17, rather than Th2 [28,37] pathway.…”

“…It is now well accepted that mesenchymal stem cells (MSCs) reside in many adult tissues [16,28,35], from which they can be easily isolated. The increasing focus on MSCs is due to the recent evidences on their possible involvement in different pathologies [32,36].…”

Amniotic Fluid-Derived Stem Cells for Cardiovascular Tissue Engineering Applications

“…27,29,30 AFSC are being explored for use in tissue engineering applications for a variety of tissue types. This review focuses on cardiovascular cell types and tissue engineering, which are addressed further below, but AFSC are also being investigated for osteogenic, 13,31,32 renal, 33,34 myogenic, 35 epithelial, 36,37 and neuronal 14,[38][39][40][41][42][43][44][45] differentiation and tissue engineering. An area of contention that remains with AFSC is whether sorting for c-kit expression is necessary or appropriate.…”

Comparative study between amniotic-fluid mesenchymal stem cells and retinal pigmented epithelium (RPE) stem cells ability to differentiate towards RPE cells

“…Human amniotic fluid samples (n°3) were obtained from amniocentesis and collected after informed consent preserving patient anonymity, as previously described (Orciani et al 2008). Briefly, 5 ml of amniotic fluid were centrifuged at 400g for 10 min and the resulting pellets were washed twice with phosphate-buffered saline (PBS; Sigma Aldrich) to remove cellular debris and blood.…”