Proinflammatory interleukins' production by adipose tissue-derived mesenchymal stromal cells: the impact of cell culture conditions and cell-to-cell interaction

2015 ◽  
Vol 33 (6) ◽  
pp. 385-392 ◽  
Author(s):  
Elena Andreeva ◽  
Irina Andrianova ◽  
Julia Rylova ◽  
Aleksandra Gornostaeva ◽  
Polina Bobyleva ◽  
...  
Keyword(s):  
Cell Culture ◽  
Adipose Tissue ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Cell Interaction ◽  
Culture Conditions ◽  
The Impact ◽  
Cell Culture Conditions

scholarly journals Exofucosylation of Adipose Mesenchymal Stromal Cells Alters Their Secretome Profile

2020 ◽  
Vol 8 ◽  
Author(s):  
David García-Bernal ◽  
Mariano García-Arranz ◽  
Ana I. García-Guillén ◽  
Ana M. García-Hernández ◽  
Miguel Blanquer ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Culture Conditions ◽  
Peripheral Tissues ◽  
Standard Culture ◽  
Anti Inflammatory ◽  
Selectin Binding ◽  
The Impact

Mesenchymal stromal cells (MSCs) constitute the cell type more frequently used in many regenerative medicine approaches due to their exclusive immunomodulatory properties, and they have been reported to mediate profound immunomodulatory effects in vivo. Nevertheless, MSCs do not express essential adhesion molecules actively involved in cell migration, a phenotypic feature that hampers their ability to home inflamed tissues following intravenous administration. In this study, we investigated whether modification by fucosylation of murine AdMSCs (mAdMSCs) creates Hematopoietic Cell E-/L-selectin Ligand, the E-selectin-binding CD44 glycoform. This cell surface glycan modification of CD44 has previously shown in preclinical studies to favor trafficking of mAdMSCs to inflamed or injured peripheral tissues. We analyzed the impact that exofucosylation could have in other innate phenotypic and functional properties of MSCs. Compared to unmodified counterparts, fucosylated mAdMSCs demonstrated higher in vitro migration, an altered secretome pattern, including increased expression and secretion of anti-inflammatory molecules, and a higher capacity to inhibit mitogen-stimulated splenocyte proliferation under standard culture conditions. Together, these findings indicate that exofucosylation could represent a suitable cell engineering strategy, not only to facilitate the in vivo MSC colonization of damaged tissues after systemic administration, but also to convert MSCs in a more potent immunomodulatory/anti-inflammatory cell therapy-based product for the treatment of a variety of autoimmune, inflammatory, and degenerative diseases.

scholarly journals Functional Characteristics of Multipotent Mesenchymal Stromal Cells from Pituitary Adenomas

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Kaspars Megnis ◽  
Ilona Mandrika ◽  
Ramona Petrovska ◽  
Janis Stukens ◽  
Vita Rovite ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Pituitary Adenomas ◽  
Cell Types ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Culture Conditions ◽  
Standard Laboratory ◽  
Vimentin Expression ◽  
Cell Culture Conditions ◽  
Supportive Cells

Pituitary adenomas are one of the most common endocrine and intracranial neoplasms. Although they are theoretically monoclonal in origin, several studies have shown that they contain different multipotent cell types that are thought to play an important role in tumor initiation, maintenance, and recurrence after therapy. In the present study, we isolated and characterized cell populations from seven pituitary somatotroph, nonhormonal, and lactotroph adenomas. The obtained cells showed characteristics of multipotent mesenchymal stromal cells as observed by cell morphology, cell surface marker CD90, CD105, CD44, and vimentin expression, as well as differentiation to osteogenic and adipogenic lineages. They are capable of growth and passaging under standard laboratory cell culture conditions and do not manifest any hormonal cell characteristics. Multipotent mesenchymal stromal cells are present in pituitary adenomas regardless of their clinical manifestation and show no considerable expression of somatostatin 1–5 and dopamine 2 receptors. Most likely obtained cells are a part of tissue-supportive cells in pituitary adenoma microenvironment.

The Hematopoietic Supportive Potential of Human Mesenchymal Stromal Cells Is Associated with Expression of Cadherins.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1402-1402
Author(s):  
Christoph Roderburg ◽  
Anke Diehlmann ◽  
Frederik Wein ◽  
Anne Faber ◽  
Ulf Krause ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Adipose Tissue ◽  
Cord Blood ◽  
Cell Line ◽  
Umbilical Cord ◽  
Mesenchymal Stromal Cells ◽  
Umbilical Cord Blood ◽  
Stromal Cells ◽  
Culture Conditions ◽  
Cells Cultured

Abstract Self renewal and differentiation of hematopoietic stem cells (HSC) are governed by interaction with the supportive microenvironment of the bone marrow. Secreted factors as well as specific cell adhesion proteins are involved in this interaction. As an in vitro model system, the hematopoietic microenvironment can be mimicked by supportive mesenchymal stromal cells (MSC). We have compared the supportive potential of human MSC from bone marrow (BM) isolated under two different culture conditions (BM-MSC-M1 and BM-MSC-M2), from adipose tissue (AT) and umbilical cord blood (CB) that were all cultivated as described before (Wagner et al. Exp Hematol.2005;11:1402–1416.). As controls we have used the human fibroblast cell line HS68 and the murine fetal liver cell line AFT024. CD34+ cells were isolated from human cord blood and cultured in direct contact with irradiated stromal cells. After four, seven and twelve days the immunophenotype of the hematopoietic cells was analyzed by flow cytometry. Many progenitor cells cultured on BM-MSC or AFT024 maintained a primitive phenotype of CD34+/CD38- cells whereas the proportion of these cells was reduced upon cultivation with CB-MSC and cells cultured on AT-MSC and HS68 displayed a significantly higher expression of CD38 and lower expression of CD34. Furthermore, long term culture initiating-cell (LTC-IC) assays were performed on the different feeder layer. LTC-IC frequency was significantly higher on BM-MSC that were isolated under the two different culture conditions (BM-MSC-M1 1,15 ±0.11%; BM-MSC-M2 1.14±0.08%) and on CB-MSC (1.10±0.13%) compared to AT-MSC (0.32±0.09%) and HS68 (0.67±0.12%). We have compared gene expression profiles of BM-MSC-M1, BM-MSC-M2, CB-MSC, AT-MSC and HS68 by cDNA microarray analysis (51,144 different cDNA clones of the RZPD3 Unigene Set). Differential expression of various genes correlated with the observed differences in supportive potential. Among these were adhesion proteins like N-cadherin, cadherin11, fibronectin1, various integrins (ITGA1, ITGA5 and ITGB1) and VCAM1 as well as secreted proteins including osteonectin, CTGF and SDF1. Westerblot analysis verified on protein level that cadherin11, N-cadherin, and ITGB1 were highly expressed on BM-MSC as compared to AT-MSC and HS68 fibroblasts. In conclusion MSC from human bone marrow or from umbilical cord blood support to a significantly higher degree the maintenance and proliferation of primitive hematopoietic progenitors than MSC derived from adipose tissue. This affinity correlated with up-regulation of cadherin11, N-cadherin and intergrin-beta1 on BM-MSC and CB-MSC.

scholarly journals Defining the mechanistic toxicity of metal oxide nanoparticles in vitro, under physiologically relevant oxygen concentrations

2021 ◽  
Author(s):  
◽  
Michael Theodoulides
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Hepg2 Cells ◽  
Metal Oxide Nanoparticles ◽  
Culture Conditions ◽  
Oxide Nanoparticles ◽  
Cell Type ◽  
Culture Environment ◽  
The Impact ◽  
Cell Culture Conditions

Metal oxide nanoparticles (MONPs) are intended for use in numerous consumer applications, leading to inevitable human exposure. Previous work conducted in hyperoxic cell culture conditions (21% O2, 5% CO2) with nanoparticles (NPs) has proven the ability of some material types to induce genotoxicity and inflammatoxicity. Alteration in intracellular calcium [i(Ca2+)] signalling is involved in facilitating toxicity through the alteration of signal-transduction pathways, but there is less understanding of the impact of NPs exposure upon changes in such signalling pathways. Furthermore, whilst human cells cultured in ambient air may induce a particular toxicity profile, this may not be the same under the physiologic oxygen conditions experienced in the human body. Therefore, the aim of this study was to assess the impact of anatase TiO2 (NM-102), Rutile TiO2 (NM-104) and dextran coated superparamagnetic Fe3O4 (dSPIONs) upon monocytes (THP-1), macrophages (dTHP-1) and hepatocarcinoma (HepG2) cells in both an in vivo-resembling physioxia environment (5%O2, 5%CO2) and hyperoxic cell culture conditions (21%O2, 5%CO2). Their impact on i(Ca2+) homeostasis and how it relates to their potential genotoxic potential was also evaluated.Due to the importance of different physicochemical characteristics for the facilitation of toxicity, all MONPs were characterized. MONPs hydrodynamic diameter (HD) and ζ-potential (ζ) in PBS were identified using dynamic light scattering: NM-102: HD=391.9nm, ζ =7.1±2.0mV; NM-104: HD=255nm, ζ=14.6 +/- 2.1mV; dSPIONs: HD=88.6nm, ζ =10.4±1.3mV. The possible toxic effect of NPs depends on their concentration and duration of their interaction with cells. Therefore, following 24h exposure to dSPIONs (0-100µg/ml), concentration-dependent and cell-type-dependent (dTHP1>THP-1>HepG2) significant increases in NP-cellular interaction were observed, which was significantly greater in the physioxic culture environment. Concurrent, significant loss of dSPION-associated cell proliferation (evaluated using relative population doubling) in all cell lines and significant increases in DNA damage was also identified in HepG2 cells (using the cytokinesis block micronucleus assay), albeit only in physioxia. Exposure to ≥10ug/ml NM-102 and NM-104 resulted in significant, two-fold increases in micronuclei formation in HepG2 in both environments. All MONPs induced a significant increase in tumour necrosis factor-α and interleukin-8 secretion in all cell lines and oxygen culture environments. Increase in the production of the chemokines was correlated with the observed HepG2 cell genotoxicity. In all cell lines and cell culture environments, treatment for up to 5h with NM-102 or dSPIONs triggered cell type specific increases in i(Ca2+) that correlated with the reduction of cellular antioxidant glutathione (measured after 5h treatment with all the MONPs). After pre-treatment of the cell lines with antioxidant trollox in all cell culture environments i(Ca2+) appeared to be increased independently from the change of cellular redox status. Environment-specific biological interaction and impacts with regard to NP uptake, genotoxic effects, and consequence on cellular signaling mechanisms were only observed with dSPIONs in a physioxic culture environment, while NM-102 and NM-104 induced similar effects in both environments. The results presented in this study allow the conclusion that the environmental oxygen content has an impact on the NP toxicity profiles although it is NP dependent.

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