Abstract
Human amniotic fluid stem cells have become an attractive stem cell source for potential applications in regenerative medicine and tissue engineering. The aim of this study was to characterize amniotic fluid-derived mesenchymal stem cells (AF-MSCs) from second- and third-trimester of gestation. Using two-stage protocol, MSCs were successfully cultured and exhibited typical stem cell morphological, specific cell surface, and pluripotency markers characteristics. AF-MSCs differentiated into adipocytes, osteocytes, chondrocytes, myocytes, and neuronal cells, as determined by morphological changes, cell staining, and RT-qPCR showing the tissue-specific gene presence for differentiated cell lineages. Using SYNAPT G2 High Definition Mass Spectrometry technique approach, we performed for the first time the comparative proteomic analysis between undifferentiated AF-MSCs from late trimester of gestation and differentiated into myogenic, adipogenic, osteogenic, and neurogenic lineages. The analysis of the functional and expression patterns of 250 high abundance proteins selected from more than 1400 demonstrated the similar proteome of cultured and differentiated AF-MSCs but the unique changes in their expression profile during cell differentiation that may help the identification of key markers in differentiated cells. Our results provide evidence that human amniotic fluid of second- and third-trimester contains stem cells with multilineage potential and may be attractive source for clinical applications.
Highlights
Human amniotic fluid (AF) collected during amniocentesis between 15th week and 19th week of gestation is used for the routine prenatal diagnosis of wide range of fetal abnormalities and genetic diseases [1,2,3,4]
Clonal populations were established with cloning rings or mechanically picked up, with immunoselection of cells expressing the receptor for Steel factor (C-kit+) or magnetic cell sorting for CD117+ [10,11,12]
We demonstrated that AFMSCs can be successfully isolated and expanded from both second- and third-trimester amniotic fluids, which maintain the expression of multipotency markers and are inducible to different cell lineages
Summary
Human amniotic fluid (AF) collected during amniocentesis between 15th week and 19th week of gestation is used for the routine prenatal diagnosis of wide range of fetal abnormalities and genetic diseases [1,2,3,4]. AF from amniocentesis samples contains terminally differentiated cells with limited proliferation capacity and fetal mesenchymal stem cells with multilineage differentiation potential [5, 6]. Multiple approaches have been used to isolate and characterize these stem cell types. Distinct clonal populations were isolated from AF by dilution and direct plating, including phenotypically and functionally distinct stromal cell clones, long-lived epithelial cells, and senescent population [9]. Clonal populations were established with cloning rings or mechanically picked up, with immunoselection of cells expressing the receptor for Steel factor (C-kit+) or magnetic cell sorting for CD117+ [10,11,12]. The majority of isolated AFSCs shared a multipotent mesenchymal phenotype and exhibited high proliferation and
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