Addressing the impact of different fetal bovine serum percentages on mesenchymal stem cells biological performance

Khasawneh, Ramada R.; Sharie, Ahmed H. Al; Rub, Ejlal Abu-El; Serhan, Abdullah Omar; Obeidat, Hayam Nizar

doi:10.1007/s11033-019-04898-1

Cited by 24 publications

(15 citation statements)

References 16 publications

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“…Recent results have also indicated the extent to which the type of serum (as an essential component for cell cultivation) and its modification are capable of affecting the biology and behavior of MSCs. Choosing a suitable FBS percentage is a controversial issue and depends merely on the experience of the researchers and the recommendations suggested by the suppliers [ 31 ]. Several differences in the sensitivity of human MSCs to various sera and their alternatives have been discussed [ 32 ].…”

Section: Discussionmentioning

confidence: 99%

In Vitro Cellular and Molecular Interplay between Human Foreskin-Derived Mesenchymal Stromal/Stem Cells and the Th17 Cell Pathway

et al. 2021

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Foreskin, considered a biological waste material, has been shown to be a reservoir of therapeutic cells. The immunomodulatory properties of mesenchymal stromal/stem cells (MSCs) from the foreskin (FSK-MSCs) are being evaluated in cell-based therapy for degenerative, inflammatory and autoimmune disorders. Within the injured/inflamed tissue, proinflammatory lymphocytes such as IL-17-producing T helper cells (Th17) may interact with the stromal microenvironment, including MSCs. In this context, MSCs may encounter different levels of T cells as well as specific inflammatory signals. Uncovering the cellular and molecular changes during this interplay is central for developing an efficient and safe immunotherapeutic tool. To this end, an in vitro human model of cocultures of FSK-MSCs and T cells was established. These cocultures were performed at different cell ratios in the presence of an inflammatory setting. After confirming that FSK-MSCs respond to ISCT criteria by showing a typical phenotype and multilineage potential, we evaluated by flow cytometry the expression of Th17 cell markers IL-17A, IL23 receptor and RORγt within the lymphocyte population. We also measured 15 human Th17 pathway-related cytokines. Regardless of the T cell/MSC ratio, we observed a significant increase in IL-17A expression associated with an increase in IL-23 receptor expression. Furthermore, we observed substantial modulation of IL-1β, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, INF-γ, sCD40, and TNF-α secretion. These findings suggest that FSK-MSCs are receptive to their environment and modulate the T cell response accordingly. The changes within the secretome of the stromal and immune environment are likely relevant for the therapeutic effect of MSCs. FSK-MSCs represent a valuable cellular product for immunotherapeutic purposes that needs to be further clarified and developed.

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Section: Discussionmentioning

confidence: 99%

In Vitro Cellular and Molecular Interplay between Human Foreskin-Derived Mesenchymal Stromal/Stem Cells and the Th17 Cell Pathway

et al. 2021

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“…Moreover, the microvesicle production and proliferation assay in Ren et al's study was performed using ultracentrifugated FBS, whereas our study used culture conditions without serum, which may explain the reduced proliferation response of the keratinocytes. The proliferation activity of MSCs depends on serum concentration, with a reduced percentage of FBS in culture media inhibiting proliferation but enhancing the immunosuppressive properties of MSCs [41]. The impact of culture conditions on the angiogenic potential of extracellular vesicles isolated from umbilical cord MSCs has been reported in studies on the biological activity of MSCs and their extracellular vesicles cultured in different types of xeno-free media [42].…”

Section: Discussionmentioning

confidence: 99%

Microvesicles from Human Immortalized Cell Lines of Endothelial Progenitor Cells and Mesenchymal Stem/Stromal Cells of Adipose Tissue Origin as Carriers of Bioactive Factors Facilitating Angiogenesis

Krawczenko

Bielawska‐Pohl

Paprocka

et al. 2020

Stem Cells International

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Endothelial progenitor cells (EPCs) and mesenchymal stem/stromal cells (MSCs) are associated with maintaining tissue homeostasis and tissue repair. Both types of cells contribute to tissue regeneration through the secretion of trophic factors (alone or in the form of microvesicles). This study investigated the isolation and biological properties of microvesicles (MVs) derived from human immortalized MSC line HATMSC1 of adipose tissue origin and EPC line. The human immortalized cell line derived from the adipose tissue of a patient with venous stasis was established in our laboratory using the hTERT and pSV402 plasmids. The human EPC line originating from cord blood (HEPC-CB.1) was established in our previous studies. Microvesicles were isolated through a sequence of centrifugations. Analysis of the protein content of both populations of microvesicles, using the Membrane-Based Antibody Array and Milliplex ELISA showed that isolated microvesicles transported growth factors and pro- and antiangiogenic factors. Analysis of the miRNA content of isolated microvesicles revealed the presence of proangiogenic miRNA (miR-126, miR-296, miR-378, and miR-210) and low expression of antiangiogenic miRNA (miR-221, miR-222, and miR-92a) using real-time RT-PCR with the TaqMan technique. The isolated microvesicles were assessed for their effect on the proliferation and proangiogenic properties of cells involved in tissue repair. It was shown that both HEPC-CB.1- and HATMSC1-derived microvesicles increased the proliferation of human endothelial cells of dermal origin and that this effect was dose-dependent. In contrast, microvesicles had a limited impact on the proliferation of fibroblasts and keratinocytes. Both types of microvesicles improved the proangiogenic properties of human dermal endothelial cells, and this effect was also dose-dependent, as shown in the Matrigel assay. These results confirm the hypothesis that microvesicles of HEPC-CB.1 and HATMSC1 origin carry proteins and miRNAs that support and facilitate angiogenic processes that are important for cutaneous tissue regeneration.

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“…EVs were successfully and reproducibly isolated from equine bone-marrow-derived MSCs using both UC and SEC, facilitating a broad comparison of UC and SEC. The MSCs were cultured in serum-free medium for 48 h to exclude possible interferences with serum during the isolation process, as EVs present in the FCS or other serum types used for cell culture can significantly affect the perceived yield of EVs and exert downstream effects, including both therapeutic as well as (xenogeneic) inflammatory effects [ 59 ].…”

Section: Discussionmentioning

confidence: 99%

“…The absence of validated antibodies is a considerable obstacle in working with equine cells. Although the horse and the human genome are highly conserved, the binding of human antibodies to equine targets is in most cases not effective [ 59 , 60 ]. BLASTED protein sequences of standard EV markers showed a homology of 84% for CD63, 92% for CD9, 98% for TSG101 and 97% for CD81 between horses and humans.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of Extracellular Vesicles from Equine Mesenchymal Stem Cells

Soukup¹,

Gerner

Gueltekin³

et al. 2022

IJMS

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Extracellular vesicles (EVs) are nanosized lipid bilayer-encapsulated particles secreted by virtually all cell types. EVs play an essential role in cellular crosstalk in health and disease. The cellular origin of EVs determines their composition and potential therapeutic effect. Mesenchymal stem/stromal cell (MSC)-derived EVs have shown a comparable therapeutic potential to their donor cells, making them a promising tool for regenerative medicine. The therapeutic application of EVs circumvents some safety concerns associated with the transplantation of viable, replicating cells and facilitates the quality-controlled production as a ready-to-go, off-the-shelf biological therapy. Recently, the International Society for Extracellular Vesicles (ISEV) suggested a set of minimal biochemical, biophysical and functional standards to define extracellular vesicles and their functions to improve standardisation in EV research. However, nonstandardised EV isolation methods and the limited availability of cross-reacting markers for most animal species restrict the application of these standards in the veterinary field and, therefore, the species comparability and standardisation of animal experiments. In this study, EVs were isolated from equine bone-marrow-derived MSCs using two different isolation methods, stepwise ultracentrifugation and size exclusion chromatography, and minimal experimental requirements for equine EVs were established and validated. Equine EVs were characterised using a nanotracking analysis, fluorescence-triggered flow cytometry, Western blot and transelectron microscopy. Based on the ISEV standards, minimal criteria for defining equine EVs are suggested as a baseline to allow the comparison of EV preparations obtained by different laboratories.

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Addressing the impact of different fetal bovine serum percentages on mesenchymal stem cells biological performance

Cited by 24 publications

References 16 publications

In Vitro Cellular and Molecular Interplay between Human Foreskin-Derived Mesenchymal Stromal/Stem Cells and the Th17 Cell Pathway

In Vitro Cellular and Molecular Interplay between Human Foreskin-Derived Mesenchymal Stromal/Stem Cells and the Th17 Cell Pathway

Microvesicles from Human Immortalized Cell Lines of Endothelial Progenitor Cells and Mesenchymal Stem/Stromal Cells of Adipose Tissue Origin as Carriers of Bioactive Factors Facilitating Angiogenesis

Characterisation of Extracellular Vesicles from Equine Mesenchymal Stem Cells

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