Mesenchymal Stem Cells (Mscs) in Scanning Electron Microscopy (SEM) World

Özen, Asuman; Sancak, İrem Gül; Tiryaki, Meral; Ceylan, Ahmet; Pınarlı, Ferda Alpaslan; Delibaşı, Tuncay

doi:10.5152/niche.2013.162

Cited by 9 publications

(6 citation statements)

References 11 publications

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“…The increase in the diameter of the cells was directly proportional to the days of the differentiation, from 2 µm in day 9 to~30 µm in day 21. This was supported by previous reports that used SEM to further confirm the differentiation of MSCs [27,28].…”

Section: Differentiation Of Imscs Into Pdgfrα-positive Cellssupporting

confidence: 86%

Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

Mussa

Khan

Srivastava

et al. 2021

IJMS

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It is evident that depletion of interstitial cells and dysfunction of nitric oxide (NO) pathways are key players in development of several gastrointestinal (GI) motility disorders such as diabetic gastroparesis (DGP). One of the main limitations of DGP research is the lack of isolation methods that are specific to interstitial cells, and therefore conducting functional studies is not feasible. The present study aims (i) to differentiate telomerase transformed mesenchymal stromal cells (iMSCs) into platelet-derived growth factor receptor-α-positive cells (PDGFRα-positive cells) using connective tissue growth factor (CTGF) and L-ascorbic acids; (ii) to investigate the effects of NO donor and inhibitor on the survival rate of differentiated PDGFRα-positive cells; and (iii) to evaluate the impact of increased glucose concentrations, mimicking diabetic hyperglycemia, on the gene expression of neuronal nitric oxide synthase (nNOS). A fibroblastic differentiation-induction medium supplemented with connective tissue growth factor was used to differentiate iMSCs into PDGFRα-positive cells. The medium was changed every day for 21 days to maintain the biological activity of the growth factors. Gene and protein expression, scanning electron and confocal microscopy, and flow cytometry analysis of several markers were conducted to confirm the differentiation process. Methyl tetrazolium cell viability, nitrite measurement assays, and immunostaining were used to investigate the effects of NO on PDGFRα-positive cells. The present study, for the first time, demonstrated the differentiation of iMSCs into PDGFRα-positive cells. The outcomes of the functional studies showed that SNAP (NO donor) increased the survival rate of differentiated PDGFRα-positive cells whereas LNNA (NO inhibitor) attenuated these effects. Further experimentations revealed that hyperglycemia produced a significant increase in expression of nNOS in PDGFRα-positive cells. Differentiation of iMSCs into PDGFRα-positive cells is a novel model to conduct functional studies and to investigate the involvement of NO pathways. This will help in identifying new therapeutic targets for treatment of DGP.

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Section: Differentiation Of Imscs Into Pdgfrα-positive Cellssupporting

confidence: 86%

Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

Mussa

Khan

Srivastava

et al. 2021

IJMS

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“…In addition, cells in P 9 maintained their spindle shape (Figure 4). These SEM results were also reported [49].…”

Section: Scanning Electron Microscope (Sem)supporting

confidence: 83%

Stromal Cell Ultrastructure

Solaiman¹

2019

Stromal Cells - Structure, Function, and Therapeutic Implications

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Regenerative medicine and tissue engineering therapies present an attractive treatment alternative to the current traditional clinical treatments. Stem cells are capable of selfrenewal and multilineage differentiation. They also have the ability to create immunomodulatory microenvironment, and thus help to minimize organ damage caused by the inflammation and cells activated by the immune system. Human bone marrow mesenchymal stem/stromal cells (MSCs) have great potential for cellular therapy, as they possess the abilities to proliferate as well as to differentiate. MSCs are present in all tissues interacting with tissue cells and easy to isolate and expand in culture. Indeed, histological examination of MSCs is one of the main goals for studying their morphology. Both the light and the electron microscopes are essential tools where the histologist can identify the structure as well as the detailed ultrastructure of these cells. This will guide users to clearly understand their behavior, both in vivo and in vitro. Thus, the aim of this chapter is to give a spot of light on these cells and their histology.

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“…Moreover, SEM showed that hMSCs adhere to PEEK with an irregular and contracted morphology that is retained also after 21 days in both OM and CM, regardless positive proliferation. Observed morphology is quite different from what displayed under conventional 2D surfaces [ 21 ], where cellular extensions and pseudopods arising from each cell and extending through each other have been reported, suggesting the contribution of the material, together with the surface architecture, to altered morphology. Notably, in OM, more resembling the in vivo presence of factors able to support bone cells and bone growth, hMSCs cultured on PEEK not only showed absence of proliferation, but a reduction in number and viability and a very poor response in terms of osteogenic commitment.…”

Section: Discussionmentioning

confidence: 61%

Superior Osteo-Inductive and Osteo-Conductive Properties of Trabecular Titanium vs. PEEK Scaffolds on Human Mesenchymal Stem Cells: A Proof of Concept for the Use of Fusion Cages

Ragni

Orfei

Bidossi

et al. 2021

IJMS

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Fusion cages composed of titanium and its alloys are emerging as valuable alternative to standard polyetheretherketone (PEEK) ones routinely used in cervical and lumbar spine surgery. Aim of this study was to evaluate osteo-inductive and osteo-conductive ability of an innovative trabecular titanium (T-Ti) scaffold on human mesenchymal stem cells (hMSCs), in both absence and presence of biochemical osteogenic stimuli. Same abilities were assessed on PEEK and standard 2D plastic surface, the latter meant as gold-standard for in vitro differentiation studies. hMSCs adhered and colonized both T-Ti and PEEK scaffolds. In absence of osteogenic factors, T-Ti triggered osteogenic induction of MSCs, as demonstrated by alkaline phosphatase activity and calcium deposition increments, while PEEK and standard 2D did not. Addition of osteogenic stimuli reinforced osteogenic differentiation of hMSCs cultured on T-Ti in a significantly higher manner with respect to standard 2D plastic culture surfaces, whereas PEEK almost completely abolished the process. T-Ti driven differentiation towards osteoblasts was confirmed by gene and marker expression analyses, even in absence of osteogenic stimuli. These results clearly indicate superior in vitro osteo-inductive and osteo-conductive capacity of T-Ti compared to PEEK, and make ground for further studies supporting the use of T-Ti cages to improve bone fusion.

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Mesenchymal Stem Cells (Mscs) in Scanning Electron Microscopy (SEM) World

Cited by 9 publications

References 11 publications

Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

Differentiated PDGFRα-Positive Cells: A Novel In-Vitro Model for Functional Studies of Neuronal Nitric Oxide Synthase

Stromal Cell Ultrastructure

Superior Osteo-Inductive and Osteo-Conductive Properties of Trabecular Titanium vs. PEEK Scaffolds on Human Mesenchymal Stem Cells: A Proof of Concept for the Use of Fusion Cages

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