Quality and freshness of human bone marrow-derived mesenchymal stem cells decrease over time after trypsinization and storage in phosphate-buffered saline

Shin, Tae Hwan; Lee, Seung‐Ah; Choi, Kyunghee; Lee, Da Yeon; Kim, Yongman; Paik, Man‐Jeong; Seo, Chan; Kang, Seok Yun; Jin, Moon Suk; Yoo, Tae Hyeon; Kang, Seong Ho; Lee, Gwang

doi:10.1038/s41598-017-01315-0

Cited by 21 publications

(47 citation statements)

References 44 publications

(50 reference statements)

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“…Although different types of MS have a high sensitivity of detection, the sample preparation process is tedious, and the selectivity for different classes of metabolites has both advantages and complications ( 43 ). In particular, metabolomic profiling of the cellular components, and target tissue metabolic reactions with gas chromatography-mass chromatography (GC/MS), without targeting a single metabolite, provides a better understanding of the biofluids, cells, and clinical conditions ( 28 , 47 – 51 ). Thus, metabolomics has been used in nanotoxicity investigations utilizing high-throughput quantitation methods ( 11 , 52 ).…”

Section: Recent Approaches For Metabolomics and Transcriptomics In Namentioning

confidence: 99%

Integration of metabolomics and transcriptomics in nanotoxicity studies

Shin

Lee

et al. 2018

BMB Rep.

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Biomedical research involving nanoparticles has produced useful products with medical applications. However, the potential toxicity of nanoparticles in biofluids, cells, tissues, and organisms is a major challenge. The ‘-omics’ analyses provide molecular profiles of multifactorial biological systems instead of focusing on a single molecule. The ‘omics’ approaches are necessary to evaluate nanotoxicity because classical methods for the detection of nanotoxicity have limited ability in detecting miniscule variations within a cell and do not accurately reflect the actual levels of nanotoxicity. In addition, the ‘omics’ approaches allow analyses of in-depth changes and compensate for the differences associated with high-throughput technologies between actual nanotoxicity and results from traditional cytotoxic evaluations. However, compared with a single omics approach, integrated omics provides precise and sensitive information by integrating complex biological conditions. Thus, these technologies contribute to extended safety evaluations of nanotoxicity and allow the accurate diagnoses of diseases far earlier than was once possible in the nanotechnology era. Here, we review a novel approach for evaluating nanotoxicity by integrating metabolomics with metabolomic profiling and transcriptomics, which is termed “metabotranscriptomics”.

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Section: Recent Approaches For Metabolomics and Transcriptomics In Namentioning

confidence: 99%

Integration of metabolomics and transcriptomics in nanotoxicity studies

Shin

Lee

et al. 2018

BMB Rep.

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“…S2c ) populations. In general, the individual populations of cells are slightly stiffer than those from the mixed population, due to the extra processing undertaken 48 .However, Fig. S2b shows that the three different subpopulations plot in different regions suggesting that there is potential to utilise size and deformability changes as a basis for separation and sorting.…”

Section: Resultsmentioning

confidence: 97%

High-throughput assessment of mechanical properties of stem cell derived red blood cells, toward cellular downstream processing

Guzniczak

Zadeh

Dempsey³

et al. 2017

Sci Rep

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Stem cell products, including manufactured red blood cells, require efficient sorting and purification methods to remove components potentially harmful for clinical application. However, standard approaches for cellular downstream processing rely on the use of specific and expensive labels (e.g. FACS or MACS). Techniques relying on inherent mechanical and physical properties of cells offer high-throughput scalable alternatives but knowledge of the mechanical phenotype is required. Here, we characterized for the first time deformability and size changes in CD34+ cells, and expelled nuclei, during their differentiation process into red blood cells at days 11, 14, 18 and 21, using Real-Time Deformability Cytometry (RT-DC) and Atomic Force Microscopy (AFM). We found significant differences (p < 0.0001; standardised mixed model) between the deformability of nucleated and enucleated cells, while they remain within the same size range. Expelled nuclei are smaller thus could be removed by size-based separation. An average Young’s elastic modulus was measured for nucleated cells, enucleated cells and nuclei (day 14) of 1.04 ± 0.47 kPa, 0.53 ± 0.12 kPa and 7.06 ± 4.07 kPa respectively. Our identification and quantification of significant differences (p < 0.0001; ANOVA) in CD34+ cells mechanical properties throughout the differentiation process could enable development of new routes for purification of manufactured red blood cells.

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“…Indeed, differences in gene expression by bone marrow-derived MSCs from different donors have been previously reported as a potential problem for their routine application in clinical practice [ 30 ]. This inter-donor variability has also been attributed to a variety of other factors, including the inherent heterogeneity of the MSC populations isolated from different individuals, the duration of their culture expansion, and the period and nature of their storage [ 31 – 33 ]. The MPCs used in the present study were all within the age range of 20–35 years, were selected on the basis of their expression of STRO-1, and were subjected to similar culture and storage conditions to minimize inter-donor cell variability.…”

Section: Discussionmentioning

confidence: 99%

Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis

Wu¹,

Shimmon

Paton

et al. 2017

Stem Cell Res Ther

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BackgroundThe pharmaceutical agent pentosan polysulfate (PPS) is known to induce proliferation and chondrogenesis of mesenchymal progenitor cells (MPCs) in vitro and in vivo. However, the mechanism(s) of action of PPS in mediating these effects remains unresolved.In the present report we address this issue by investigating the binding and uptake of PPS by MPCs and monitoring gene expression and proteoglycan biosynthesis before and after the cells had been exposed to limited concentrations of PPS and then re-established in culture in the absence of the drug (MPC priming).MethodsImmuno-selected STRO-1+ mesenchymal progenitor stem cells (MPCs) were prepared from human bone marrow aspirates and established in culture. The kinetics of uptake, shedding, and internalization of PPS by MPCs was determined by monitoring the concentration-dependent loss of PPS media concentrations using an enzyme-linked immunosorbent assay (ELISA) and the uptake of fluorescein isothiocyanate (FITC)-labelled PPS by MPCs. The proliferation of MPCs, following pre-incubation and removal of PPS (priming), was assessed using the Wst-8 assay method, and proteoglycan synthesis was determined by the incorporation of 35SO4 into their sulphated glycosaminoglycans. The changes in expression of MPC-related cell surface antigens of non-primed and PPS-primed MPCs from three donors was determined using flow cytometry. RNA sequencing of RNA isolated from non-primed and PPS-primed MPCs from the same donors was undertaken to identify the genes altered by the PPS priming protocol.ResultsThe kinetic studies indicated that, in culture, PPS rapidly binds to MPC surface receptors, followed by internalisation and localization within the nucleus of the cells. Following PPS-priming of MPCs and a further 48 h of culture, both cell proliferation and proteoglycan synthesis were enhanced. Reduced expression of MPC-related cell surface antigen expression was promoted by the PPS priming, and RNA sequencing analysis revealed changes in the expression of 42 genes.ConclusionThis study has shown that priming of MPCs with low concentrations of PPS enhanced chondrogenesis and MPC proliferation by modifying their characteristic basal gene and protein expression. These findings offer a novel approach to re-programming mesenchymal stem cells for clinical indications which require the repair or regeneration of cartilaginous tissues such as in osteoarthritis and degenerative disc disease.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0723-y) contains supplementary material, which is available to authorized users.

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Quality and freshness of human bone marrow-derived mesenchymal stem cells decrease over time after trypsinization and storage in phosphate-buffered saline

Cited by 21 publications

References 44 publications

Integration of metabolomics and transcriptomics in nanotoxicity studies

Integration of metabolomics and transcriptomics in nanotoxicity studies

High-throughput assessment of mechanical properties of stem cell derived red blood cells, toward cellular downstream processing

Pentosan polysulfate binds to STRO-1+ mesenchymal progenitor cells, is internalized, and modifies gene expression: a novel approach of pre-programing stem cells for therapeutic application requiring their chondrogenesis

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