scholarly journals Optimal Pore Size of Honeycomb Polylactic Acid Films for In Vitro Cartilage Formation by Synovial Mesenchymal Stem Cells

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Misaki Yagi ◽  
Mitsuru Mizuno ◽  
Ryota Fujisawa ◽  
Hisako Katano ◽  
Kentaro Endo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Pore Size ◽  
Polylactic Acid ◽  
Time Lapse ◽  
Induction Medium ◽  
High Porosity ◽  
Cartilage Formation ◽  
Selection Of

Background. Tissue engineering of cartilage requires the selection of an appropriate artificial scaffold. Polylactic acid (PLA) honeycomb films are expected to be highly biodegradable and cell adhesive due to their high porosity. The purpose of this study was to determine the optimal pore size of honeycomb PLA films for in vitro cartilage formation using synovial mesenchymal stem cells (MSCs). Methods. Suspensions of human synovial MSCs were plated on PLA films with different pore sizes (no pores, or with 5 μm or 20 μm pores) and then observed by scanning electron microscopy. The numbers of cells remaining in the film and passing through the film were quantified. One day after plating, the medium was switched to chondrogenic induction medium, and the films were time-lapse imaged and observed histologically. Results. The 5 μm pore film showed MSCs with pseudopodia that extended between several pores, while the 20 μm pore film showed MSC bodies submerged into the pores. The number of adhered MSCs was significantly lower for the film without pores, while the number of MSCs that passed through the film was significantly higher for the 20 μm pore film. MSCs that were induced to form cartilage peeled off as a sheet from the poreless film after one day. MSCs formed thicker cartilage at two weeks when growing on the 5 μm pore films than on the 20 μm pore films. Conclusions. Honeycomb PLA films with 5 μm pores were suitable for in vitro cartilage formation by synovial MSCs.

scholarly journals Positively Correlated CD47 Activation and Autophagy in Umbilical Cord Blood-Derived Mesenchymal Stem Cells during Senescence

2021 ◽  
Vol 2021 ◽  
pp. 1-13
Author(s):  
Gee-Hye Kim ◽  
Yun Kyung Bae ◽  
Ji Hye Kwon ◽  
Miyeon Kim ◽  
Soo Jin Choi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Growth ◽  
Cell Therapy ◽  
Critical Role ◽  
Therapeutic Effects ◽  
Stem Cell Maintenance ◽  
Selection Of

Autophagy plays a critical role in stem cell maintenance and is related to cell growth and cellular senescence. It is important to find a quality-control marker for predicting senescent cells. This study verified that CD47 could be a candidate to select efficient mesenchymal stem cells (MSCs) to enhance the therapeutic effects of stem cell therapy by analyzing the antibody surface array. CD47 expression was significantly decreased during the expansion of MSCs in vitro ( p < 0.01 ), with decreased CD47 expression correlated with accelerated senescence phenotype, which affected cell growth. UCB-MSCs transfected with CD47 siRNA significantly triggered the downregulation of pRB and upregulation of pp38, which are senescence-related markers. Additionally, autophagy-related markers, ATG5, ATG12, Beclin1, and LC3B, revealed significant downregulation with CD47 siRNA transfection. Furthermore, autophagy flux following treatment with an autophagy inducer, rapamycin, has shown that CD47 is a key player in autophagy and senescence to maintain and regulate the growth of MSCs, suggesting that CD47 may be a critical key marker for the selection of effective stem cells in cell therapy.

Morphological examination during in vitro cartilage formation by human mesenchymal stem cells

2005 ◽  
Vol 322 (2) ◽  
pp. 217-226 ◽  
Author(s):  
Shizuko Ichinose ◽  
Motoki Tagami ◽  
Takeshi Muneta ◽  
Ichiro Sekiya
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Human Mesenchymal Stem Cells ◽  
Morphological Examination ◽  
Cartilage Formation

scholarly journals The interplay between chondrocyte spheroids and mesenchymal stem cells boosts cartilage regeneration within a 3D natural-based hydrogel

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Annachiara Scalzone ◽  
Ana M. Ferreira ◽  
Chiara Tonda-Turo ◽  
Gianluca Ciardelli ◽  
Kenny Dalgarno ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Sol Gel ◽  
Cartilage Regeneration ◽  
High Porosity ◽  
Young’S Moduli ◽  
Chondrogenic Phenotype ◽  
Sol Gel Transition ◽  
Gel Transition

Abstract Articular cartilage (AC) lacks the ability to self-repair and cell-based approaches, primarily based on using chondrocytes and mesenchymal stem cells (MSCs), are emerging as effective technology to restore cartilage functionality, because cells synergic functionality may support the maintenance of chondrogenic phenotype and promote extracellular matrix regeneration. This work aims to develop a more physiologically representative co-culture system to investigate the influence of MSCs on the activity of chondrocytes. A thermo-sensitive chitosan-based hydrogel, ionically crosslinked with β–glycerophosphate, is optimised to obtain sol/gel transition at physiological conditions within 5 minutes, high porosity with pores diameter <30 µm, and in vitro mechanical integrity with compressive and equilibrium Young’s moduli of 37 kPa and 17 kPa, respectively. Live/dead staining showed that after 1 and 3 days in culture, the encapsulated MSCs into the hydrogels are viable and characterised by round-like morphology. Furthermore chondrocyte spheroids, seeded on top of gels that contained either MSCs or no cells, show that the encapsulated MSCs stimulate chondrocyte activity within a gel co-culture, both in terms of maintaining the coherence of chondrocyte spheroids, leading to a larger quantity of CD44 (by immunofluorescence) and a higher production of collagen and glycosaminoglycans (by histology) compared with the mono-culture.

Stro-1 Positive and Stro-1− Negtive Human Mesenchymal Stem Cells Express Different Levels of Immunosuppression.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4964-4964
Author(s):  
Dominique Thierry 9 ◽  
Y. Z. Zhang 1 ◽  
A. Chapel 2 ◽  
M. Benshidoum 3 ◽  
C. Mazurier 4 ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Lymphocyte Proliferation ◽  
Human Mesenchymal Stem Cells ◽  
Clinical Use ◽  
Soluble Factors ◽  
The Difference ◽  
Tgf Β1 ◽  
Selection Of

Abstract Mesenchymal stem cells (MSCs), have been shown to elicit immunosuppressive effect on allogeneic lymphocyte response. However, MSCs are heterogeneous and data on the inhibitory abilities of different MSC subsets are lacking. In the present study, we selected Stro-1+ cells from human bone marrow and evaluated the inhibitory capability of this MSC subset in mixed lymphocyte reactions (MLRs) or in mitogen stimulation asssays, in comparison to that of Stro-1− cells. To evaluate the two MSC subsets for immunomodulation in vitro, we added 1,000–30,000 Stro-1+ or Stro-1− cells to MLR at the beginning of the experiment. When comparing the inhibitory effects of the two subsets, PBLs proliferation was significantly more inhibited by Stro-1+MSCs (11.0%–63.7%) than by stro-1−MSCs (35.5%-106%) (P<0.01). Furthermore, as few as 1,000 Stro-1+ MSC could inhibit lymphocyte proliferation more effectively than 10 times more (10,000 cells) Stro-1−cells. As it was observed with the mixed lymphocyte reaction, suppression of the response to the mitogen also occurred in a dose dependent fashion, but to a lesser extent with the Stro-1−cells (25.5%–80.1% vs 7.5%–38.4% in Stro-1+cells) (P<0.05). To investigate whether the difference of suppressive effect that we observed between Stro-1+ and Stro-1− cells, still exist when MSC subsets are separated physically from PBL, we performed MLR in the upper chamber of a transwell and we seeded the lower chamber either with Stro-1+ or Stro-1− cells. The inhibitory effect of Stro-1+ cells was significantly more profound than the one observed when Stro-1− cells were used in the Transwell culture system (p<0.05) (Figure 3), demonstrating that one or several soluble factors was involved in production of different suppressive effects. Cytokine and chemokine genes, IL-10, TGF-β1, SDF-1, SCF and IL-6 expression were evaluated in both MSC subsets by quantitative RT-PCR. Low levels of IL-6, SCF, SDF-1 were observed in Stro-1+, which induced a fold increase around 1 (0,96 ± 0,32; 0,96 ± 0,24; 0,96 ± 0,24), indicating that there is no signifiant difference of these genes expression between the two MSC subsets. However, we observed in Stro-1+ a decreased gene expression for IL-10 (0,24 fold ± 0,59; p <0,05) and for TGF b1 (0,43 fold ± 0,32; p <0,05). This finding suggested that the candidate T-cell inhibitory factors TGF b1, IL-10, which are lower expressed in Stro-1+ cells, are not responsible for the more profound inhibition of immunoreactivity by Stro-1+ cells. We show here that significant differences do exist within these two subsets. Stro-1+ cells inhibit lymphocyte proliferation significantly more profoundly than Stro-1−cells. The difference is in part mediated by soluble factors, but not IL-10 and TGF-β1. These results point to the notion that Stro-1+ cells can elicit more powerful immunosuppressive ability and a pre-selection of Stro-1+MSC for clinical use may be advisable. These findings suggest that pre-selection of MSC before clinical use might produce more effective immunosuppression in different therapeutic applications, especially in clinics for the prevention of graft versus host disease (GVHD).

scholarly journals Neuron-Specific Fluorescence Reporter-Based Live Cell Tracing for Transdifferentiation of Mesenchymal Stem Cells into Neurons by Chemical Compound

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Do Won Hwang ◽  
Hyun Woo Kwon ◽  
Jaeho Jang ◽  
Hee Jung Jung ◽  
Kwang Rok Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Neuronal Differentiation ◽  
Small Molecule ◽  
Plasmid Vector ◽  
Time Lapse ◽  
Live Cell ◽  
Easy Method ◽  
Specific Fluorescence

Although transdifferentiation of mesenchymal stem cells (MSCs) into neurons increases the possibility of therapeutic use of MSCs for neurodevelopmental disorders, the use of MSCs has the limitation on differentiation efficiency to neuronal lineage and lack of an easy method to monitor the transdifferentiation. In this study, using time-lapse live cell imaging, we assessed the neuronal differentiation of MSCs induced by a small molecule “NHPDQC (N-hydroxy-2-oxo-3-(3-phenylprophyl)-1,2-dihydroquinoxaline-6-carboxamide, C18H17N3O3).” Plasmid vector containing red fluorescence reporter genes under the control of the tubulin α1 (Tα1) promoter (pTα1-DsRed2) traced the neuronal differentiation of MSCs. Two days after NHPDQC treatment, MSCs showed neuron-like phenotype with neurite outgrowth and high expression of neuron-specific markers in more than 95% cells. The fluorescence signals increased in the cytoplasm of pTα1-DsRed2-transfected MSCs after NHPDQC treatment. In vitro monitoring of MSCs along the time courses showed progressive increase of fluorescence till 30 h after treatment, corresponding with the increase in neurite length. We examined an efficient neuronal differentiation of MSCs by NHPDQC alone and monitored the temporal changes of neuronal differentiation by neuron-specific fluorescence reporter along time. This method would help further our understanding of the differentiation of MSCs to produce neurons by simple treatment of small molecule.

scholarly journals Petaloid recombinant peptide enhances in vitro cartilage formation by synovial mesenchymal stem cells

2018 ◽  
Vol 37 (6) ◽  
pp. 1350-1357 ◽  
Author(s):  
Mana Naritomi ◽  
Mitsuru Mizuno ◽  
Hisako Katano ◽  
Nobutake Ozeki ◽  
Koji Otabe ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Recombinant Peptide ◽  
Cartilage Formation

In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel

2005 ◽  
Vol 322 (2) ◽  
pp. 289-298 ◽  
Author(s):  
Akiko Yokoyama ◽  
Ichiro Sekiya ◽  
Kyosuke Miyazaki ◽  
Shizuko Ichinose ◽  
Yuiro Hata ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Collagen Gel ◽  
Cartilage Formation

scholarly journals Polydopamine-based biofunctional substrate coating promotes mesenchymal stem cell migration

MRS Advances ◽  
2021 ◽  
Author(s):  
Zijun Deng ◽  
Weiwei Wang ◽  
Xun Xu ◽  
Nan Ma ◽  
Andreas Lendlein
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Migration ◽  
Time Lapse ◽  
Implant Surface ◽  
Tissue Integration ◽  
Coated Substrate ◽  
Fibronectin Deposition

Abstract Rapid migration of mesenchymal stem cells (MSCs) on device surfaces could support in vivo tissue integration and might facilitate in vitro organoid formation. Here, polydopamine (PDA) is explored as a biofunctional coating to effectively promote MSC motility. It is hypothesized that PDA stimulates fibronectin deposition and in this way enhances integrin-mediated migration capability. The random and directional cell migration was investigated by time-lapse microscopy and gap closure assay respectively, and analysed with softwares as computational tools. A higher amount of deposited fibronectin was observed on PDA substrate, compared to the non-coated substrate. The integrin β1 activation and focal adhesion kinase (FAK) phosphorylation at Y397 were enhanced on PDA substrate, but the F-actin cytoskeleton was not altered, suggesting MSC migration on PDA was regulated by integrin initiated FAK signalling. This study strengthens the biofunctionality of PDA coating for regulating stem cells and offering a way of facilitating tissue integration of devices. Graphic abstract Polydopamine-coated substrate induces increased fibronectin deposition of mesenchymal stem cells, and promotes cell migration via integrin-initiated FAK signaling, compared to non-coated polystyrene-based standard tissue culture surface. In this way, multifunctional PDA coating could support in vivo tissue integration on implant surface and promote in vitro organoid formation.

scholarly journals Selection of DNA Aptamers for Differentiation of Human Adipose-Derived Mesenchymal Stem Cells from Fibroblasts

2021 ◽  
Author(s):  
Mariane Izabella Abreu de Melo ◽  
Pricila da Silva Cunha ◽  
Marcelo Coutinho de Miranda ◽  
Joana Lobato Barbosa ◽  
Jerusa Araújo Quintão Arantes Faria ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dissociation Constants ◽  
Binding Capacity ◽  
Cell Types ◽  
Dermal Fibroblasts ◽  
Cell Therapies ◽  
Promising Tool ◽  
Selection Of

Abstract In recent years, stem cell therapy has shown promise in regenerative medicine. In this context, the distinction of mesenchymal stem cells from fibroblasts has been crucial for safe clinical application of these cells. In the present study, we developed aptamers capable of specifically recognize mesenchymal stem cells derived from adipose tissue (ASC) using the Cell-SELEX technique. We tested the affinity of ASC aptamers compared to dermal fibroblasts (FIB). Quantitative PCR was advantageous for the validation of four candidate aptamers in vitro. The binding capabilities of Apta 2 and Apta 42 could not distinguish both cell types, while Apta 21 and Apta 99 showed a better binding capacity to ASC with dissociation constants (Kd) of 50.46 ± 2.28 nM and 72.71 ± 10.3 nM, respectively. However, Apta 21 showed a Kd of 86.78 ± 9.14 nM when incubated with FIB. Therefore, only Apta 99 showed specificity to detect ASC. This aptamer is a promising tool for the in vitro identification of ASC. These results will help understand the differences between these two cell types for more specific and precise cell therapies.

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