Comparative proteomic analysis of two distinct stem-cell populations from human amniotic fluid

Abstract

Human amniotic fluid (AF) contains a variety of stem cells of embryonic and extra-embryonic origins.We characterized two distinct types of stem cells isolated from residual AF material derived from prenatal diagnostic amniocentesis. The two types of cells differed in their morphology and growth kinetics, showing fast (fast human amniotic stem cells; fHASCs) or slow (slow human amniotic stem cells; sHASCs) population-doubling times. Both fHASCs and sHASCs expressed pluripotent stem-cell markers, yet unlike sHASCs, clonogenic fHASCs would generate embryoid bodies and maintain their original phenotype during prolonged in vitro passaging. fHASCs -but not sHASCs -expressed the KLF4, SSEA-4 and CD117 markers. Differential proteomic analysis allowed us to identify the protein patterns specific for either cell type as potentially contributing to their distinct phenotypes. We found thirty-six proteins that were differentially expressed by the two cell types, and those proteins were classified according to their biological and molecular functions. Bioinformatic cluster analysis revealed differential occurrence of cytoskeletal proteins, such as vimentin, F-actin-binding protein, and chloride intracellular channel protein 1. Selected proteins differentially expressed by fHASCs and sHASCs were further characterized by Western blot analysis and confocal microscopy.

High Quality Independent From a Donor: Human Amniotic Fluid Derived Stem Cells—A Practical Analysis Based on 165 Clinical Cases

“…The presented results confirmed that hAFSCs have potential to differentiate toward adipocyte, chondrocyte, and osteoblast cell line, which is consistent with the literature data [De Coppi et al, 2007;Saulnier et al, 2009;You et al, 2009;Phermthai et al, 2010;DeKoninck et al, 2014;Sousa et al, 2014;Romani et al, 2015].…”

“…Those cells secrete a wide variety of growth factors with complex feedback mechanisms among many types of immune cells [Murphy et al, ]. Recent discoveries of stem cells population in amniotic fluid have postulated that the well characterized amniotic fluid is a promising alternative source of fetal stem cells for therapy [Pappa and Anagnou, ; You et al, ; Da Sacco et al, ; Klemmt et al, ; Cananzi and De Coppi, ; Chun et al, ; Piskorska‐Jasiulewicz and Witkowska‐Zimny, ; Romani et al, ; Si et al, ]. It was observed during this study that within the primary culture, there are cells with different morphology.…”

The Glyoxalase System Is a Novel Cargo of Amniotic Fluid Stem-Cell-Derived Extracellular Vesicles

“…Written informed consent was obtained from each woman. HASCs were isolated and cultured as previously described [ 26 , 27 , 28 ]. Briefly, fresh amniotic fluid (3–5 mL) was centrifuged and the cell pellet cultured in 18% Chang B/2% Chang C media (Irvine Scientific, Ireland, UK) for 6–7 days to allow adherent cells to form colonies.…”

“…Cells with a predominantly fibroblast-like morphology and a colony shape similar to dermatoglyphics, appropriately characterized, represented HASCs and were cultured in MSCGM medium (Lonza, Gaithersburg, MD, USA). Since HASCs characterization in all the cases showed similarity in all the parameters considered for their characterization [ 26 , 27 , 28 ], only one HASC cell line was considered to recover EVs.…”

Effect of the Enrichment in c-Kit Stem Cell Potential of Foetal Human Amniotic Fluid Cells: Characterization from Single Cell Analysis to the Secretome Content

“…Even looking only at hAFSCs obtained during the second trimester (from amniocentesis), various classifications have been proposed for human amniotic subpopulations, including a morphological classification, since up to three cell types have been described, from epithelioid to spindle-shaped [6]. Therefore, amniocytes differ in many functional aspects, including survival rate, plastic adhesion, and growth characteristics.…”