Molecular and Phenotypic Characterization of Human Amniotic Fluid-Derived Cells: A Morphological and Proteomic Approach

Pipino, Caterina; PierdomenicoLaura,; TomoPamela, Di; GiuseppeFabrizio, Di; CianciEleonora,; D'AlimonteIolanda,; Morabito, Caterina; Centurione, Lucia; AntonucciIvana,; Mariggiò, Maria Addolorata; PietroRoberta, Di; Ciccarelli, Renata; Marchisio, Marco; Romano, Mario; AngelucciStefania,; Pandolfi, Assunta

doi:10.1089/scd.2014.0453

Cited by 30 publications

(33 citation statements)

References 53 publications

(31 reference statements)

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“…Although no differences in expression of MSC-associated surface markers were observed between the young and elderly MSC populations (Additional file 1 : Figure S1), SSEA-4, originally identified on embryonic stem cells and a characteristic of actively dividing MSCs [ 15 , 25 , 26 ], was found to be expressed by a smaller fraction of the elderly vs young MSCs (Fig. 1i ).…”

Section: Resultsmentioning

confidence: 93%

Restoring the quantity and quality of elderly human mesenchymal stem cells for autologous cell-based therapies

et al. 2017

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BackgroundDegenerative diseases are a major public health concern for the aging population and mesenchymal stem cells (MSCs) have great potential for treating many of these diseases. However, the quantity and quality of MSCs declines with aging, limiting the potential efficacy of autologous MSCs for treating the elderly population.MethodsHuman bone marrow (BM)-derived MSCs from young and elderly donors were obtained and characterized using standard cell surface marker criteria (CD73, CD90, CD105) as recommended by the International Society for Cellular Therapy (ISCT). The elderly MSC population was isolated into four subpopulations based on size and stage-specific embryonic antigen-4 (SSEA-4) expression using fluorescence-activated cell sorting (FACS), and subpopulations were compared to the unfractionated young and elderly MSCs using assays that evaluate MSC proliferation, quality, morphology, intracellular reactive oxygen species, β-galactosidase expression, and adenosine triphosphate (ATP) content.ResultsThe ISCT-recommended cell surface markers failed to detect any differences between young and elderly MSCs. Here, we report that elderly MSCs were larger in size and displayed substantially higher concentrations of intracellular reactive oxygen species and β-galactosidase expression and lower amounts of ATP and SSEA-4 expression. Based on these findings, cell size and SSEA-4 expression were used to separate the elderly MSCs into four subpopulations by FACS. The original populations (young and elderly MSCs), as well as the four subpopulations, were then characterized before and after culture on tissue culture plastic and BM-derived extracellular matrix (BM-ECM). The small SSEA-4-positive subpopulation representing ~ 8% of the original elderly MSC population exhibited a “youthful” phenotype that was similar to that of young MSCs. The biological activity of this elderly subpopulation was inhibited by senescence-associated factors produced by the unfractionated parent population. After these “youthful” cells were isolated and expanded (three passages) on a “young microenvironment” (i.e., BM-ECM produced by BM cells from young donors), the number of cells increased ≈ 17,000-fold to 3 × 109 cells and retained their “youthful” phenotype.ConclusionsThese results suggest that it is feasible to obtain large numbers of high-quality autologous MSCs from the elderly population and establish personal stem cell banks that will allow serial infusions of “rejuvenated” MSCs for treating age-related diseases.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0688-x) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 93%

Restoring the quantity and quality of elderly human mesenchymal stem cells for autologous cell-based therapies

et al. 2017

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“…with changes of medium every 3 days. To visualize calcium sediments, cultures were stained with Alizarin Red S (Sigma-Aldrich) and the microscope images were acquired (Pipino et al, 2015).…”

Section: Flow Cytometrymentioning

confidence: 99%

Mesenchymal stromal cells from amniotic fluid are less prone to senescence compared to those obtained from bone marrow: An in vitro study

Alessio

Pipino

Mandatori

et al. 2018

Journal Cellular Physiology

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Mesenchymal stromal cells (MSCs) are considered to be an excellent source in regenerative medicine. They contain several cell subtypes, including multipotent stem cells. MSCs are of particular interest as they are currently being tested using cell and gene therapies for a number of human diseases. They represent a rare population in tissues; for this reason, they require, before being transplanted, an in vitro amplification. This process may induce replicative senescence, thus affecting differentiation and proliferative capacities. Increasing evidence suggests that MSCs from fetal tissues are significantly more plastic and grow faster than MSCs from bone marrow. Here, we compare amniotic fluid mesenchymal stromal cells (AF-MSCs) and bone marrow mesenchymal stromal cells (BM-MSCs) in terms of cell proliferation, surface markers, multidifferentiation potential, senescence, and DNA repair capacity. Our study shows that AF-MSCs are less prone to senescence with respect to BM-MSCs. Moreover, both cell models activate the same repair system after DNA damage, but AF-MSCs are able to return to the basal condition more efficiently with respect to BM-MSCs. Indeed, AF-MSCs are better able to cope with genotoxic stress that may occur either during in vitro cultivation or following transplantation in patients. Our findings suggest that AF-MSCs may represent a valid alternative to BM-MSCs in regenerative medicine, and, of great relevance, the investigation of the mechanisms involved in DNA repair capacity of both AF-MSCs and BM-MSCs may pave the way to their rational use in the medical field.

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“…Moreover, a dim expression of the embryonic stem antigen SSEA-4 is observed. To refine the analysis, the SSEA-4 stained cells were permeabilized to detect also the intracellular levels of the molecule 26,27) . As expected, the cells are negative for the leukocyte antigen CD45 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Osteoblastic differentiating potential of dental pulp stem cells <i>in vitro</i> cultured on a chemically modified microrough titanium surface

et al. 2018

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Titanium surface modification is critical for dental implant success. Our aim was to determine surfaces influence on dental pulp stem cells (DPSCs) viability and differentiation. Implants were divided into sandblasted/acid-etched (control) and sandblasted/acid-etched coated with calcium and magnesium ions (CaMg), supplied as composite, (test). Proliferation was evaluated by MTT, differentiation checking osteoblastic gene expression, PGE2 secretion and matrix formation, inflammation by Interleukin 6 (IL-6) detection. MTT and IL-6 do not modify on test. A PGE2 increase on test is recorded. BMP2 is higher on test at early experimental points, Osterix and RUNX2 augment later. Alizarin-red S reveals higher matrix production on test. These results suggest that test surface is more osteoinductive, representing a start point for in vivo studies aiming at the construction of more biocompatible dental implants, whose integration and clinical performance are improved and some undesired effects, such as implant stability loss and further surgical procedures, are reduced.

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Molecular and Phenotypic Characterization of Human Amniotic Fluid-Derived Cells: A Morphological and Proteomic Approach

Cited by 30 publications

References 53 publications

Restoring the quantity and quality of elderly human mesenchymal stem cells for autologous cell-based therapies

Restoring the quantity and quality of elderly human mesenchymal stem cells for autologous cell-based therapies

Mesenchymal stromal cells from amniotic fluid are less prone to senescence compared to those obtained from bone marrow: An in vitro study

Osteoblastic differentiating potential of dental pulp stem cells <i>in vitro</i> cultured on a chemically modified microrough titanium surface

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