Human Amniotic Fluid Mesenchymal Stem Cells from Second- and Third-Trimester Amniocentesis: Differentiation Potential, Molecular Signature, and Proteome Analysis

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.

Characteristics of Full-Term Amniotic Fluid-Derived Mesenchymal Stem Cells in Different Culture Media

“…The hAFMSCs cultured in DMEM-FBS moderately expressed CD105 and CD166. A similar expression pattern of CD105 and CD166 was also observed by other researchers during generation of MSCs derived from second-and third-trimester amniotic fluid using FBS-supplemented media [13,16,18]. CD105 plays a role in chondrogenic differentiation [27], while CD166 is involved in neurite extension [28].…”

“…However, in DMEM-FBS, only about 50% of the P3 cultured cells expressed CD105 and CD166. Lower percentage of cells expressing CD105 and CD166 was also reported by several researchers when second-and third-trimester hAFMSCs were cultured in different types of media supplemented with FBS [13,16,18]. The proliferation of the hAF cells is slower in DMEM-FBS than in SFM.…”

“…At this gestational stage, it is impossible to collect a larger volume of amniotic fluid, and there are increased risks of uterine contamination and miscarriage [13]. Alternatively, stem cells could be isolated from amniotic fluid of full-term pregnancies, specifically during delivery.…”

Hematopoietic & Mesenchymal - The Two Lineages of Bone Marrow Stem Cells

“…Adipose tissue is one of the richest sources of MSC's. Adipose stem cells are currently actively being researched in clinical trials for treatment in a variety of diseases [16,17].…”

Adipogenic and Osteogenic Markers Characterization of Human Amniotic Fluid Stem Cells

“…These cells have the capability in treatment of diseases such as diabetes, bone defects, neural degeneration, blood disorders and heart and kidney disease [7] through its capabilities of differentiating into adipocytes, osteocytes, chondrocytes, myocytes, and neuronal cells [8].…”