Surface functionalization of tissue culture polystyrene plates with hydroxyapatite under body fluid conditions and its effect on differentiation behaviors of mesenchymal stem cells

The benefits of combined cell/material therapy appear promising for myocardial infarction treatment. The safety of alginate, along with its excellent biocompatibility and biodegradability, has been extensively investigated for cardiac tissue engineering. Among graphene-based nanomaterials, reduced graphene oxide has been considered as a promising candidate for cardiac treatment due to its unique physicochemical properties. In this study, the reduced graphene oxide incorporation effect within alginate hydrogels was investigated for cardiac repair application. Reduced graphene oxide reinforced alginate properties, resulting in an increase in gel stiffness. The cytocompatibility of the hydrogels prepared with human bone marrow–derived mesenchymal stem cells was assessed by the 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide) assay. Following reduced graphene oxide addition, alginate-reduced graphene oxide retained significantly higher cell viability compared to that of alginate and cells cultured on tissue culture plates. Acridine orange/propidium iodide staining was also used to identify both viable and necrotic human bone marrow–derived mesenchymal stem cells within the prepared hydrogels. After a 72-h culture, the percentage of viable cells was twice as much as those cultured on either alginate or tissue culture plate, reaching approximately 80%. Quantitative reverse transcription polymerase chain reaction analysis was performed to assess gene expression of neonatal rat cardiac cells encapsulated on hydrogels for TrpT-2, Conx43, and Actn4 after 7 days. The expression of all genes in alginate-reduced graphene oxide increased significantly compared to that in alginate or tissue culture plate. The results obtained confirmed that the presence of reduced graphene oxide, as an electro-active moiety within alginate, could tune the physicochemical properties of this material, providing a desirable electroactive hydrogel for stem cell therapy in patients with ischemic heart disease.

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3D Culture of MSCs on a Gelatin Microsphere in a Dynamic Culture System Enhances Chondrogenesis

International Journal of Molecular Sciences ◽

10.3390/ijms21082688 ◽

2020 ◽

Vol 21 (8) ◽

pp. 2688

Author(s):

Shamsul Sulaiman ◽

Shiplu Roy Chowdhury ◽

Mh Busra Fauzi ◽

Rizal Abdul Rani ◽

Nor Hamdan Mohamad Yahaya ◽

...

Keyword(s):

Stem Cells ◽

Tissue Culture ◽

Mesenchymal Stem Cells ◽

Chondrogenic Differentiation ◽

3D Culture ◽

Mechanical Agitation ◽

Dynamic Culture ◽

Culture Plate ◽

Gelatin Microsphere ◽

Tissue Culture Plate

Recent advancement in cartilage tissue engineering has explored the potential of 3D culture to mimic the in vivo environment of human cartilaginous tissue. Three-dimensional culture using microspheres was described to play a role in driving the differentiation of mesenchymal stem cells to chondrocyte lineage. However, factors such as mechanical agitation on cell chondrogenesis during culture on the microspheres has yet to be elucidated. In this study, we compared the 2D and 3D culture of bone-marrow-derived mesenchymal stem cells (BMSCs) on gelatin microspheres (GMs) in terms of MSC stemness properties, immune-phenotype, multilineage differentiation properties, and proliferation rate. Then, to study the effect of mechanical agitation on chondrogenic differentiation in 3D culture, we cultured BMSCs on GM (BMSCs-GM) in either static or dynamic bioreactor system with two different mediums, i.e., F12: DMEM (1:1) + 10% FBS (FD) and chondrogenic induction medium (CIM). Our results show that BMSCs attached to the GM surface and remained viable in 3D culture. BMSCs-GM proliferated faster and displayed higher stemness properties than BMSCs on a tissue culture plate (BMSCs-TCP). GMs also enhanced the efficiency of in-vitro chondrogenesis of BMSCs, especially in a dynamic culture with higher cell proliferation, RNA expression, and protein expression compared to that in a static culture. To conclude, our results indicate that the 3D culture of BMSCs on gelatin microsphere was superior to 2D culture on a standard tissue culture plate. Furthermore, culturing BMSCs on GM in dynamic culture conditions enhanced their chondrogenic differentiation.

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Improvement of in situ Follicular Activation and Early Development in Cryopreserved Human Ovarian Cortical Tissue by Co-Culturing with Mesenchymal Stem Cells

Cells Tissues Organs ◽

10.1159/000506303 ◽

2019 ◽

Vol 208 (1-2) ◽

pp. 48-58

Author(s):

Marzieh Hosseini ◽

Saghar Salehpour ◽

Marefat Ghaffari Novin ◽

Zahra Shams Mofarahe ◽

Mohammad-Amin Abdollahifar ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Cell Proliferation ◽

Tissue Culture ◽

Mesenchymal Stem Cells ◽

Ovarian Tissue ◽

Follicular Development ◽

Cortical Tissue ◽

Culture Systems ◽

The Impact

Follicular loss and tissue degeneration are great challenges in ovarian tissue culture systems. Mesenchymal stem cells (MSC) secrete a cocktail of growth factors and cytokines which supports adjacent cells and tissues. The aim of the current study was to investigate the impact of human bone marrow (hBM)-MSC, as co-culture cells, on human follicular development in ovarian cortical tissue (OCT) culture. For this purpose, warmed OCT fragments were co-cultured with hBM-MSC for 8 days and compared to monocultured OCT. During the culture period, ovarian follicle survival and development in the OCT were evaluated using histological observation, follicular developmental-related genes expression, and estradiol production. Furthermore, cell proliferation and apoptosis were assessed. The results showed that there were no significant differences in conserved ovarian follicles with a normal morphology between the two groups. However, the percentage of developing follicles, as well as follicular developmental gene expression, significantly increased in the co-culture group compared to the monoculture group. On the other hand, compared with the monoculture group, the co-culture group demonstrated a significant increase in cell proliferation, indicated by Ki67 gene expression, as well as a dramatic decrease in apoptotic cell percentage, revealed by TUNEL assay. These findings indicated that co-culturing of hBM-MSC with OCT could improve follicular activation and early follicular development in human ovarian tissue culture systems.

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Laminin-511 and recombinant vitronectin supplementation enables human pluripotent stem cell culture and differentiation on conventional tissue culture polystyrene surfaces in xeno-free conditions

Journal of Materials Chemistry B ◽

10.1039/d1tb01878g ◽

2021 ◽

Author(s):

Ya-Chu Liu ◽

Lee-Kiat Ban ◽

Henry Hsin-Chung Lee ◽

Hsin-Ting Lee ◽

Yu-Tang Chang ◽

...

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Extracellular Matrix ◽

Tissue Culture ◽

Pluripotent Stem Cells ◽

Pluripotent Stem Cell ◽

Culture Conditions ◽

Human Pluripotent Stem Cell ◽

Tissue Culture Polystyrene ◽

Ecm Proteins

Human pluripotent stem cells (hPSCs) are typically cultivated on extracellular matrix (ECM) protein-coated dishes in xeno-free culture conditions. We supplemented mixed ECM proteins (laminin-511 and recombinant vitronectin, rVT) in culture...

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Production of Adult Human Synovial Fluid-Derived Mesenchymal Stem Cells in Stirred-Suspension Culture

Stem Cells International ◽

10.1155/2018/8431053 ◽

2018 ◽

Vol 2018 ◽

pp. 1-16 ◽

Cited By ~ 6

Author(s):

Kristen D. Jorgenson ◽

David A. Hart ◽

Roman Krawetz ◽

Arindom Sen

Keyword(s):

Stem Cells ◽

Tissue Culture ◽

Mesenchymal Stem Cells ◽

Synovial Fluid ◽

Cell Attachment ◽

Operating Cost ◽

Cartilage Regeneration ◽

Fold Increase ◽

Microcarrier Beads ◽

Cytodex 3

The chondrogenic potential of synovial fluid-derived mesenchymal stem cells (SF-MSCs) supports their use in cartilage regeneration strategies. However, their paucity in synovial fluid necessitates their proliferation in culture to generate clinically relevant quantities. Here it was determined that 125 mL stirred suspension bioreactors utilizing Cytodex-3 microcarrier beads represent a viable platform for the proliferation of these cells. During the inoculation phase, a bead loading of 2 g/L, an inoculation ratio of 4.5 cells/bead, and continuous agitation at 40 rpm in a medium with 5% serum resulted in high cell attachment efficiencies and a subsequent overall cell fold expansion of 5.7 over 8 days. During the subsequent growth phase, periodic addition of new microcarriers and fresh medium increased culture longevity, resulting in a 21.3 cell fold increase over 18 days in the same vessel without compromising the defining characteristics of the cells. Compared to static tissue culture flasks, a bioreactor-based bioprocess requires fewer handling steps, is more readily scalable, and for the same cell production level, has a lower operating cost as it uses approximately half the medium. Therefore, stirred suspension bioreactors incorporating microcarrier technology represent a viable and more efficient platform than tissue culture flasks for the generation of SF-MSCs in culture.

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Improved Isolation of Mesenchymal Stem Cells Based on Interactions between N-Acetylglucosamine-Bearing Polymers and Cell-Surface Vimentin

Stem Cells International ◽

10.1155/2019/4341286 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 2

Author(s):

Hirohiko Ise ◽

Kumiko Matsunaga ◽

Marie Shinohara ◽

Yasuyuki Sakai

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Tissue Culture ◽

Mesenchymal Stem Cells ◽

Bone Marrow Cells ◽

Cell Types ◽

Binding Activity ◽

Isolation System ◽

Multiple Cell

Mesenchymal stem cells (MSCs) in bone marrow and adipose tissues are expected to be effective tools for regenerative medicine to treat various diseases. To obtain MSCs that possess both high differentiation and tissue regenerative potential, it is necessary to establish an isolation system that does not require long-term culture. It has previously been reported that the cytoskeletal protein vimentin, expressed on the surfaces of multiple cell types, possesses N-acetylglucosamine- (GlcNAc-) binding activity. Therefore, we tried to exploit this interaction to efficiently isolate MSCs from rat bone marrow cells using GlcNAc-bearing polymer-coated dishes. Cells isolated by this method were identified as MSCs because they were CD34-, CD45-, and CD11b/c-negative and CD90-, CD29-, CD44-, CD54-, CD73-, and CD105-positive. Osteoblast, adipocyte, and chondrocyte differentiation was observed in these cells. In total, yields of rat MSCs were threefold to fourfold higher using GlcNAc-bearing polymer-coated dishes than yields using conventional tissue-culture dishes. Interestingly, MSCs isolated with GlcNAc-bearing polymer-coated dishes strongly expressed CD106, whereas those isolated with conventional tissue-culture dishes had low CD106 expression. Moreover, senescence-associated β-galactosidase activity in MSCs from GlcNAc-bearing polymer-coated dishes was lower than that in MSCs from tissue-culture dishes. These results establish an improved isolation method for high-quality MSCs.

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Response of mouse bone marrow mesenchymal stem cells to graphene-containing grid-like bioactive glass scaffolds produced by robocasting

Journal of Biomaterials Applications ◽

10.1177/0885328218799610 ◽

2018 ◽

Vol 33 (4) ◽

pp. 488-500

Author(s):

Aylin M Deliormanlı ◽

Mert Türk ◽

Harika Atmaca

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Simulated Body Fluid ◽

Bioactive Glass ◽

Body Fluid ◽

Composite Scaffolds ◽

Toxic Response ◽

Marrow Mesenchymal Stem Cells

In the study, three-dimensional, grid-like silicate-based bioactive glass scaffolds were manufactured using a robotic deposition technique. Inks were prepared by mixing 13-93 bioactive glass particles in Pluronic® F-127 solution. After deposition, scaffolds were dried at room temperature and sintered at 690°C for 1 h. The surface of the sintered scaffolds was coated with graphene nanopowder (1, 3, 5, 10 wt%) containing poly(ε-caprolactone) solution. The in vitro mineralization ability of the prepared composite scaffolds was investigated in simulated body fluid. The surface of the simulated body fluid-treated scaffolds was analyzed using scanning electron microscopy to investigate the hydroxyapatite formation. Mechanical properties were tested under compression. Results revealed that graphene coating has no detrimental effect on the hydroxyapatite forming ability of the prepared glass scaffolds. On the other hand, it decreased the compression strength of the scaffolds at high graphene concentrations. The prepared grid-like bioactive glass-based composite scaffolds did not show toxic response to bone marrow mesenchymal stem cells. It was shown that stem cells seeded onto the scaffolds attached and proliferated well on the surface. Cells seeded on the scaffolds surface also demonstrated osteogenic differentiation under in vitro conditions in the absence of transforming growth factors.

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