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Isolation and molecular characterization of amniotic fluid-derived mesenchymal stem cells obtained from Caesarean sections

ABSTRACT: Human amniotic fluid cells (AFCs) are immune-privileged with low immunogenicity and anti-inflammatory properties. Furthermore, they are high proliferative and have a broad differentiation potential, making them amenable for cell replacement therapies. Amniotic fluid (AF) is routinely obtained via amniocentesis. It contains heterogeneous populations of foetal-derived differentiated and undifferentiated progenitor cells including mesenchymal stem cells (MSCs). AF-derived mesenchymal stem cells (AF-MSCs) can self-renew and have a high proliferation and differentiation potential. In this study, we isolated AFCs from AF obtained during Caesarean sections (C-sections) and characterized them. The AFCs showed typical MSCs characteristics in relation to morphology, in vitro differentiation potential, cell surface marker expression and secreted factors. Subpopulations of AF-MSCs expressed several pluripotency-associated markers such as stage specific embryonic antigen 4 (SSEA4), c-Kit, TRA-1-60 and TRA-1-81, making them reprogrammable to induced pluripotent stem cells (iPSCs) without the use of ectopic gene expression. Additionally, they express the mesenchymal marker - Vimentin and multipotency-associated stem cell marker - CD133. Furthermore, the transcriptome and secretome analyses showed significant overlap with bone marrow-derived MSCs. C-section-derived AFMSCs can be routinely obtained without any risk to the foetus. Patient-specific AF-MSCs can be used for personalized cell therapies and disease modelling.

ORGANISM(S): Homo sapiens

PROVIDER: GSE100448 | GEO | 2018/01/12

SECONDARY ACCESSION(S): PRJNA391824

REPOSITORIES: GEO

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Project description:The raw files of the proteomics dataset are N=18 and a brief description of the files is shown below: AF1 raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 1 AF2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 2 AFA1.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 3 AFA2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 4 AFB1.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 5 AFB2.raw: exosomes derived from Amniotic Fluid-Mesenchymal Stromal cells 6 HL1A.raw: exosomes derived from Hepatocyte like cells 1 HL1B.raw: exosomes derived from Hepatocyte like cells 2 HL2A.raw: exosomes derived from Hepatocyte like cells 3 HL2B.raw: exosomes derived from Hepatocyte like cells 4 HL3A.raw: exosomes derived from Hepatocyte like cells 5 HL3B.raw: exosomes derived from Hepatocyte like cells 6 HPLA1.raw: exosomes derived from Hepatic Progenitor-like cells 1 HPLA2.raw: exosomes derived from Hepatic Progenitor-like cells 2 HPLB1.raw: exosomes derived from Hepatic Progenitor-like cells 3 HPLB2.raw: exosomes derived from Hepatic Progenitor-like cells 4 HPLA.raw: exosomes derived from Hepatic Progenitor-like cells 5 HPLB.raw: exosomes derived from Hepatic Progenitor-like cells 6 Raw files analysis was performed with Proteome Discoverer 1.4 (Thermo) software package, using the Sequest search engine and the Uniprot human (Homo sapiens) reviewed database, downloaded on December 15, 2017, including 20,243 entries. The search was performed using carbamidomethylation of cysteine as static and oxidation of methionine as dynamic modifications. Two missed cleavage sites, a precursor mass tolerance of 10 ppm and fragment mass tolerance of 0.05 Da were allowed. False discovery rate (FDR) validation was based on q value: target FDR : 0.05. Label free quantification was performed by utilizing the precursor ion area values exported from the total ion chromatogram as defined by the Proteome Discoverer v. 1.4.0.288 (Thermo Scientific). Output files from Proteome Discoverer were processed with R programming language for statistical computing (version 4.0.3). The following comparisons were made: AF vs HL, AF vs HPL and HL vs HPL. The msf files are N=18 and have the same name as the raw files above.

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Isolation and molecular characterization of amniotic fluid-derived mesenchymal stem cells obtained from Caesarean sections

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