Characteristics Of Mesenchymal Stem Cells Research Articles

Cannabidiol Rescues TNF-α-Inhibited Proliferation, Migration, and Osteogenic/Odontogenic Differentiation of Dental Pulp Stem Cells.

Strategies to promote dental pulp stem cells (DPSCs) functions including proliferation, migration, pro-angiogenic effects, and odontogenic/osteogenic differentiation are in urgent need to restore pulpitis-damaged dentin/pulp regeneration and DPSCs-based bone tissue engineering applications. Cannabidiol (CBD), an active component of Cannabis sativa has shown anti-inflammation, chemotactic, anti-microbial, and tissue regenerative potentials. Based on these facts, this study aimed to analyze the effect of CBD on DPSCs proliferation, migration, and osteogenic/odontogenic differentiation in basal and inflammatory conditions. Highly pure DPSCs with characteristics of mesenchymal stem cells (MSCs) were successfully isolated, as indicated by the results of flowcytometry and multi-lineage (osteogenic, adipogenic, and chondrogenic) differentiation potentials. Among the concentration tested (0.1-12.5 µM), CBD (2.5 μM) showed the highest anabolic effect on the proliferation and osteogenic/odontogenic differentiation of DPSCs. Pro-angiogenic growth factor VEGF mRNA expression was robustly higher in CBD-treated DPSCs. CBD also prompted the migration of DPSCs and CBD receptor CB1 and CB2 expression in DPSCs. TNF-α inhibited the viability, migration, and osteogenic/odontogenic differentiation of DPSCs and CBD reversed these effects. CBD alleviated the TNF-α-upregulated expression of pro-inflammatory cytokines TNF-α, interleukin (IL)-1β, and IL-6 in DPSCs. In conclusion, our results indicate the possible application of CBD on DPSCs-based dentin/pulp and bone regeneration.

Whole transcriptomic analysis of mesenchymal stem cells cultured in Nichoid micro-scaffolds.

Mesenchymal stem cells (MSCs) are known to be ideal candidates for clinical applications where not only regenerative potential but also immunomodulation ability is fundamental. Over the last years, increasing efforts have been put into the design and fabrication of 3D synthetic niches, conceived to emulate the native tissue microenvironment and aiming at efficiently controlling the MSC phenotype in vitro. In this panorama, our group patented an engineered microstructured scaffold, called Nichoid. It is fabricated through two-photon polymerization, a technique enabling the creation of 3D structures with control of scaffold geometry at the cell level and spatial resolution beyond the diffraction limit, down to 100nm. The Nichoid's capacity to maintain higher levels of stemness as compared to 2D substrates, with no need for adding exogenous soluble factors, has already been demonstrated in MSCs, neural precursors, and murine embryonic stem cells. In this work, we evaluated how three-dimensionality can influence the whole gene expression profile in rat MSCs. Our results show that at only 4days from cell seeding, gene activation is affected in a significant way, since 654 genes appear to be differentially expressed (392 upregulated and 262 downregulated) between cells cultured in 3D Nichoids and in 2D controls. The functional enrichment analysis shows that differentially expressed genes are mainly enriched in pathways related to the actin cytoskeleton, extracellular matrix (ECM), and, in particular, cell adhesion molecules (CAMs), thus confirming the important role of cell morphology and adhesions in determining the MSC phenotype. In conclusion, our results suggest that the Nichoid, thanks to its exclusive architecture and 3D cell adhesion properties, is not only a useful tool for governing cell stemness but could also be a means for controlling immune-related MSC features specifically involved in cell migration.

Sequential passage and characterization of mesenchymal stem cells derived from single and pooled human umbilical cord tissue

Sequential passage and characterization of mesenchymal stem cells derived from single and pooled human umbilical cord tissue

Mesenchymal Stem Cell Therapy in Kidney Diseases: Potential and Challenges.

Kidney disease (KD) is a life-threatening disease characterized by high morbidity and mortality in clinical settings, which can be caused by many reasons, and the incidence increases with age. However, supportive therapy and kidney transplantation still have limitations in alleviating KD progression. Recently, mesenchymal stem cells (MSCs) have shown great potential in repairing injury through their multidirectional differentiation and self-renewal ability. Of note, MSCs serve as a safe and effective therapeutic strategy for treating KD in preclinical and clinical trials. Functionally, MSCs ameliorate KD progression by regulating the immune response, renal tubular cell apoptosis, tubular epithelial-mesenchymal transition, oxidative stress, angiogenesis, and so on. In addition, MSCs exhibit remarkable efficacy in both acute kidney injury (AKI) and chronic kidney disease (CKD) through paracrine mechanisms. In this review, we outline the biological characteristics of MSCs, discuss the efficacy and mechanisms of MSCs-based therapy for KD, summarize the completed and ongoing clinical trials, as well as analyze limitations and new strategies, aiming to provide new ideas and approaches for the preclinical experiments and clinical trials of MSCs transplantation for KD.

Adipose and lipoma stem cells: A donor-matched comparison.

Lipomas are slow growing benign fat tumors that develop in soft tissues of the mesoderm. Thus, the specific (dys-)function of mesenchymal stem cells (MSCs) has been suggested in the development of lipomas, but details of the tumor pathogenesis remain unclear. Existing studies comparing stem cells from native adipose (adipose stem cells [ASCs]) and lipomatous tissues (LSCs) have reported contradicting findings. However, harvesting ASCs and LSCs from different individuals might have influenced proper comparison. Therefore, we aimed to characterize donor-matched ASCs and LSCs to investigate metabolic activity, proliferation, capability for tri-linear differentiation (chondrogenesis, adipogenesis, osteogenesis), and the secretome of mature adipocytes and lipomacytes. Both stem cell types did not differ in metabolic activity, but ASCs demonstrated stronger proliferation than LSCs. While there was no difference in proteoglycan accumulation during chondrogenic differentiation, adipogenesis was higher in ASCs, with more lipid vacuole formation. Conversely, LSCs showed increased osteogenesis by higher calcium deposition. Lipomacytes showed stronger secretory activity and released higher levels of certain adipokines. Our findings indicated that LSCs possessed important characteristics of MSCs, including ASCs. However, LSCs' low proliferation and adipogenic differentiation behavior did not appear to account for enhanced tissue proliferation, but the secretome of lipomacytes could contribute to lipomatous neoplasm.

Periodontal ligament cells from patients with treated stable periodontitis: Characterization and osteogenic differentiationpotential.

Periodontal ligament progenitor cells (PDL cells) isolated from patients with inflammatory periodontitis have impaired regenerative capacity, but it is unknown whether this capacity can be recovered upon treatment and stabilization of the periodontal condition. The study aimed to investigate the expression of surface markers and the proliferation and osteogenic potential of PDL cells isolated from patients with treated stable periodontitis (S-PDL cells), periodontally healthy individuals (H-PDL cells), and patients with inflammatory periodontitis (I-PDL cells). H-PDL, I-PDL, and S-PDL cells were isolated from the extracted teeth of individuals who (1) were periodontally healthy, (2) had inflammatory periodontitis, and (3) had treated stable periodontitis, respectively. The expression levels of surface markers and the proliferative and osteogenic capacities of the PDL cells were assessed. PDL cells derived from all three sources exhibited mesenchymal stem cell (MSC) characteristics. They were positive for MSC-related markers and negative for a hematopoiesis-related marker. However, S-PDL cells had higher proliferation rates, higher expression levels of osteogenic markers, higher alkaline phosphatase activity, and more calcium nodules than I-PDL cells. But all of these parameters remained lower in S-PDL cells than in H-PDL cells. S-PDL cells proliferated faster and had greater osteogenic potential than I-PDL cells, although these values remained lower than those in H-PDL cells.

Current problems and persprectives of the treatment of cartilage injuries using mesenchimal stem cells

Annotation. Since standard methods of treatment of cartilage trauma and inflammatory diseases lack effectiveness, at present one of the most promising strategies is mesenchymal stem cell transplantation. Despite the successes in this field, many issues, such as the selection of the tissue-source of mesenchymal stem cells (MSCs) and their optimization or modification in culture in order to achieve the maximum therapeutic effect, remains an actual task for both global and domestic scientists. The purpose of the study is to prepare a review of literature sources for the period 2004-2022 (in the vast majority on publications of the last five years) and to systematize new data on the biological characteristics of MSCs from various tissue sources, which are used in strategies for the treatment of cartilage lesions tissues, to analyze existing problems and further prospects, as well as potential ways of optimizing MSC cultivation methods to increase their regenerative potential. A study of 49 sources that met the inclusion criteria was carried out. A significant variety of strategies for the use of MSCs in animal models and in clinical practice was revealed. The main mechanisms ensuring the therapeutic effect of MSCs have been determined and can be used as criteria for optimizing cellular material. However, it was found that the question of the effectiveness of the use of differentiated MSCs, the selection of the tissue-source of MSCs, and the specifics of the use of MSCs from neonatal sources remains open. Thus, the analysis of literary sources revealed further promising ways to improve the therapeutic efficiency of MSC grafting in a wide range of cartilaginous lesions. There is a need to research the effects of MSCs from neonatal tissues, and especially the umbilical cord. This will become possible due to experimental research that will be carried out in the future.

Therapeutic Potential of Mesenchymal Stem Cells in the Treatment of Epilepsy and Their Interaction with Antiseizure Medications

Epilepsy is a life-threatening neurological disease that affects approximately 70 million people worldwide. Although the vast majority of patients may be successfully managed with currently used antiseizure medication (ASM), the search for alternative therapies is still necessary due to pharmacoresistance in about 30% of patients with epilepsy. Here, we review the effects of ASMs on stem cell treatment when they could be, as expected, co-administered. Indeed, it has been reported that ASMs produce significant effects on the differentiation and determination of stem cell fate. In addition, we discuss more recent findings on mesenchymal stem cells (MSCs) in pre-clinical and clinical investigations. In this regard, their ability to differentiate into various cell types, reach damaged tissues and produce and release biologically active molecules with immunomodulatory/anti-inflammatory and regenerative properties make them a high-potential therapeutic tool to address neuroinflammation in different neurological disorders, including epilepsy. Overall, the characteristics of MSCs to be genetically engineered, in order to replace dysfunctional elements with the aim of restoring normal tissue functioning, suggested that these cells could be good candidates for the treatment of epilepsy refractory to ASMs. Further research is required to understand the potential of stem cell treatment in epileptic patients and its interaction with ASMs.

Single-Cell and Spatial Transcriptomics Decodes Wharton's Jelly-Derived Mesenchymal Stem Cells Heterogeneity and a Subpopulation with Wound Repair Signatures.

The highly heterogeneous characteristics of Wharton's jelly mesenchymal stem cells (WJ-MSCs) may be responsible for the poor clinical outcomes and poor reproducibility of treatments based on WJ-MSCs. Exploration of WJ-MSC heterogeneity with multimodal single-cell technologies will aid in establishing accurate MSC subtyping and developing screening protocols for dominant functional subpopulations. Here, the characteristics of WJ-MSCs are systematically analyzed by single cell and spatial transcriptome sequencing. Single-cell transcriptomics analysis identifies four WJ-MSC subpopulations, namely proliferative_MSCs, niche-supporting_MSCs, metabolism-related_MSCs and biofunctional-type_MSCs. Furthermore, the transcriptome, cellular heterogeneity, and cell-state trajectories of these subpopulations are characterized. Intriguingly, the biofunctional-type MSCs (marked by S100A9, CD29, and CD142) selected in this study exhibit promising wound repair properties in vitro and in vivo. Finally, by integrating omics data, it has been found that the S100A9+ CD29+ CD142+ subpopulation is more enriched in the fetal segment of the umbilical cord, suggesting that this subpopulation deriving from the fetal segment may have potential for developing into an ideal therapeutic agent for wound healing. Overall, the presented study comprehensively maps the heterogeneity of WJ-MSCs and provides an essential resource for future development of WJ-MSC-based drugs.

Free Fatty Acids from Type 2 Diabetes Mellitus Serum Remodel Mesenchymal Stem Cell Lipids, Hindering Differentiation into Primordial Germ Cells.

Type 2 diabetes mellitus (T2DM) adversely affects the essential characteristics of adipose tissue-derived mesenchymal stem cells (AdMSCs). Given that T2DM is associated with an altered serum free fatty acid (FFA) profile, we examined whether diabetic serum FFAs influence the viability, differentiation, and fatty acid composition of the major lipid fractions of human AdMSCs in vitro. Serum FFAs were isolated from 7 diabetic and 10 healthy nondiabetic female individuals. AdMSCs were cultured and differentiated into primordial germ cell-like cells (PGCLCs) in the presence of either diabetic or nondiabetic FFAs. Cell viability was assessed using trypan blue staining. Cell differentiation was evaluated by measuring the PGCLC transcriptional markers Blimp1 and Stella. Lipid fractionation and fatty acid quantification were performed using thin-layer chromatography and gas-liquid chromatography, respectively. Both diabetic and nondiabetic FFAs significantly reduced the viability of PGCLCs. The gene expression of both differentiation markers was significantly lower in cells exposed to diabetic FFAs than in those treated with nondiabetic FFAs. Saturated fatty acids were significantly increased and linoleic acid was significantly decreased in the cellular phospholipid fraction after exposure to diabetic FFAs. In contrast, monounsaturated fatty acids were reduced and linoleic acid was elevated in the cellular triglyceride fraction in response to diabetic FFAs. Such an altered serum FFA profile in patients with T2DM reduces the proliferation and differentiation potential of AdMSCs, presumably due to the aberrant distribution of fatty acids into cell phospholipids and triglycerides.

Characteristics of mesenchymal stem cells from supracrestal gingival connective tissue.

Although periodontal ligament-derived mesenchymal stem cells (PDL-MSCs) have been recognized as the best MSC choice for periodontal regeneration, using PDL-MSCs requires tooth extraction for cell isolation. The supracrestal gingival (SG) connective tissue is a part of the gingiva which is located close to the PDL. SG-MSCs might have similar characteristic to the PDL-MSCs and serve as a good MSC candidate for periodontal regeneration. This study aimed to investigate and compare the characteristics of human MSCs isolated from SG tissue (hSG-MSCs), marginal gingival tissue (hMG-MSCs), and PDL (hPDL-MSCs) in terms of MSCs properties and differential gene expression profile. Human periodontal tissue from five healthy subjects, including SG, MG, and PDL, was harvested. The primary cells of the hSG-MSCs, hMG-MSCs, and hPDL-MSCs were isolated and expanded to assess MSCs markers by flow cytometry, colony-forming ability, differentiation potential, RNA sequencing, and real-time polymerase chain reaction (PCR). Of the three cell types, the hSG-MSCs demonstrated the highest colony-forming ability. The number of alizarin red S positive colonies produced by the hSG-MSCs was higher than the hMG-MSCs but lower than the hPDL-MSCs. RNA sequencing revealed that the hSG-MSCs had a more similar gene expression profile to the hPDL-MSCs than the hMG-MSCs. Pathway enrichment analysis found no significant differences in the differentially expressed genes between the hSG-MSCs and hPDL-MSCs; however, there were significantly enriched pathways between the hPDL-MSCs and hMG-MSCs for the extracellular matrix (ECM) organization and ECM-receptor interaction pathways. This study demonstrated a close relationship between hSG-MSCs and hPDL-MSCs. hSG-MSCs could be a potential MSC source for periodontal tissue engineering.

Fabrication of 3D oriented carbon nanofiber by two-nuzzle electrospinning as a cell scaffold.

One of the important issues in tissue engineering has been the development of 3D scaffolds, which guide cells to grow functional tissues and allow the diffusion of nutrients, metabolites, and soluble factors. Factors governing scaffold design include considerations of pore size and morphology, mechanical properties versus porosity, surface properties, and appropriate biodegradability. Three-dimensional structures with low density, high surface area and porosity can be utilized effectively in the tissue engineering. Recently two-nozzle electrospinning was used for fabricate polymeric and ceramic bulky layers with specific formulation. Fabrication of 3D carbon nanofiber with this method was investigated in this assay with FESEM, TGA-DTA, FTIR and XRD. Polyacrylonitrile was used as precursor. The collector speed was changed (15, 30, 60, 150, 300 and 450rpm) to result in oriented 3D carbon nanofiber after stepwise thermal process under neutral gas atmosphere. The effect of the mechanical force applied by the collector rotation not only can arranged carbon fiber mat but also can change the crystallinity of the carbon structure. The viability and growth capability of cells on nanofibers towards the lowest cytotoxicity of them proved by MTT test. The growth characteristic of neural and mouse bone marrow mesenchymal stem cells cultured in the webs showed the good adhesion with the blown web relative to a normal electrospun mat. The electrospun nanofibers mat had good tensile properties and high porosity and provides a favorable environment for neural cell attachment and proliferation comparable to other scaffolds. The cell viability and cell growth capability in prepared nanofibers were assessed.

Comparison of the yields and properties of dedifferentiated fat cells and mesenchymal stem cells derived from infrapatellar fat pads.

Infrapatellar fat pad (IFP)-derived mesenchymal stem cells (MSCs) have high chondrogenic potential and are attractive cell sources for cartilage regeneration. During ceiling culture to acquire the characteristics of MSCs, mature adipocytes from fat tissue are known to undergo dedifferentiation, generating dedifferentiated fat (DFAT) cells. The purpose of the present study was to compare the yields and biological properties of IFP-derived MSCs and IFP-derived DFAT cells. IFPs were harvested from the knees of 8 osteoarthritis (OA) patients. DFAT cells were obtained using a ceiling culture of adipocytes isolated from the floating top layer of IFP digestion. MSCs were obtained by culturing precipitated stromal vascular fraction cells. We compared the P0 cell yields, surface antigen profile, colony formation ability, and multipotency of DFAT cells and MSCs. The P0 cell yields per flask and the estimated total cell yields from 1g of IFP were much greater for MSCs than for DFAT cells. Both MSCs and DFAT cells were positive for MSC markers. No obvious difference was observed in colony formation ability. In differentiation assays, DFAT cells produced greater amounts of lipid droplets, calcified tissue, and glycosaminoglycan than MSCs did. Adipogenic and chondrogenic gene expressions were upregulated in DFAT cells. IFP-derived DFAT cells showed higher adipogenic and chondrogenic potentials than IFP-derived MSCs, but they had a poor cell yield.

Investigation of the Apoptotic Effects of Mesenchymal Stem Cells on KG-1 Leukemic Cell Line

Background: Mesenchymal stem cells (MSCs) are undifferentiated cells with the ability of multi-potency, pluripotency, and self-renewal. MSCs show great promise in cancer therapy due to their unique features. MSC secrete various cytokines with multifunctional properties, although their roles are unclear. Methods: We have investigated the influence of secreted cytokines from MSCs on KG-1 cells as a cell model of acute myeloid leukemia (AML). For this purpose, following the culture and characterization of MSCs, a trans-well system was used for co-culturing MSCs and KG-1. To determine apoptosis induction Ki/Caspase-3 assay was conducted for cultured KG-1 alone and in co-culture with MSCs (10:1) on day 7. In the following step, the protein was isolated from both groups (control and experimental) and western blotting was done for investigating the BAX and BCL-2 proteins expression. Results: It was found that MSCs significantly enhanced caspase-3 activity in KG-1 cells (P<0.05). Besides, A significant increase in protein expression of BAX was detected, while BCL-2 displayed a dramatic reduction (P<0.01). Conclusion: As a concluding remark, MSCs have a contributory role in the apoptosis of KG-1 cells that is mediated by Caspase-3, BAX, and BCL2 expression.

The role of mesenchymal stem cell transplantation for ischemic stroke and recent research developments.

Ischemic stroke is a common cerebrovascular disease that seriously affects human health. However, most patients do not practice self-care and cannot rely on the current clinical treatment for guaranteed functional recovery. Stem cell transplantation is an emerging treatment studied in various central nervous system diseases. More importantly, animal studies show that transplantation of mesenchymal stem cells (MSCs) can alleviate neurological deficits and bring hope to patients suffering from ischemic stroke. This paper reviews the biological characteristics of MSCs and discusses the mechanism and progression of MSC transplantation to provide new therapeutic directions for ischemic stroke.

Osteogenic differentiation and proliferation potentials of human bone marrow and umbilical cord-derived mesenchymal stem cells on the 3D-printed hydroxyapatite scaffolds

Mesenchymal stem cells (MSCs) are a promising candidate for bone repair. However, the maintenance of MSCs injected into the bone injury site remains inefficient. A potential approach is to develop a bone-liked platform that incorporates MSCs into a biocompatible 3D scaffold to facilitate bone grafting into the desired location. Bone tissue engineering is a multistep process that requires optimizing several variables, including the source of cells, osteogenic stimulation factors, and scaffold properties. This study aims to evaluate the proliferation and osteogenic differentiation potentials of MSCs cultured on 2 types of 3D-printed hydroxyapatite, including a 3D-printed HA and biomimetic calcium phosphate-coated 3D-printed HA. MSCs from bone marrow (BM-MSCs) and umbilical cord (UC-MSCs) were cultured on the 3D-printed HA and coated 3D-printed HA. Scanning electron microscopy and immunofluorescence staining were used to examine the characteristics and the attachment of MSCs to the scaffolds. Additionally, the cell proliferation was monitored, and the ability of cells to differentiate into osteoblast was assessed using alkaline phosphatase (ALP) activity and osteogenic gene expression. The BM-MSCs and UC-MSCs attached to a plastic culture plate with a spindle-shaped morphology exhibited an immunophenotype consistent with the characteristics of MSCs. Both MSC types could attach and survive on the 3D-printed HA and coated 3D-printed HA scaffolds. The MSCs cultured on these scaffolds displayed sufficient osteoblastic differentiation capacity, as evidenced by increased ALP activity and the expression of osteogenic genes and proteins compared to the control. Interestingly, MSCs grown on coated 3D-printed HA exhibited a higher ALP activity and osteogenic gene expression than those cultured on the 3D-printed HA. The finding indicated that BM-MSCs and UC-MSCs cultured on the 3D-printed HA and coated 3D-printed HA scaffolds could proliferate and differentiate into osteoblasts. Thus, the HA scaffolds could provide a suitable and favorable environment for the 3D culture of MSCs in bone tissue engineering. Additionally, biomimetic coating with octacalcium phosphate may improve the biocompatibility of the bone regeneration scaffold.

Cryopreservation does not change the performance and characteristics of allogenic mesenchymal stem cells highly over-expressing a cytoplasmic therapeutic transgene for cancer treatment

BackgroundMesenchymal stem cells (MSCs) driven gene directed enzyme prodrug therapy is a promising approach to deliver therapeutic agents to target heterogenous solid tumours. To democratize such a therapy, cryopreservation along with cold chain transportation is an essential part of the logistical process and supply chain. Previously, we have successfully engineered MSCs by a non-viral DNA transfection approach for prolonged and exceptionally high expression of the fused transgene cytosine deaminase, uracil phosphoribosyl transferase and green fluorescent protein (CD::UPRT::GFP). The aim of this study was to determine the effects of cryopreservation of MSCs engineered to highly overexpress this cytoplasmic therapeutic transgene.MethodsModified MSCs were preserved in a commercially available, GMP-grade cryopreservative—CryoStor10 (CS10) for up to 11 months. Performance of frozen-modified MSCs was compared to freshly modified equivalents in vitro. Cancer killing potency was evaluated using four different cancer cell lines. Migratory potential was assessed using matrigel invasion assay and flow cytometric analysis for CXCR4 expression. Frozen-modified MSC was used to treat canine patients via intra-tumoral injections, or by intravenous infusion followed by a daily dose of 5-flucytosine (5FC).ResultsWe found that cryopreservation did not affect the transgene expression, cell viability, adhesion, phenotypic profile, and migration of gene modified canine adipose tissue derived MSCs. In the presence of 5FC, the thawed and freshly modified MSCs showed comparable cytotoxicity towards one canine and three human cancer cell lines in vitro. These cryopreserved cells were stored for about a year and then used to treat no-option-left canine patients with two different types of cancers and notably, the patients showed progression-free interval of more than 20 months, evidence of the effectiveness in treating spontaneously occurring cancers.ConclusionThis study supports the use of cryopreserved, off-the-shelf transiently transfected MSCs for cancer treatment.

Mesenchymal Stem Cell Secreted-Extracellular Vesicles are Involved in Chondrocyte Production and Reduce Adipogenesis during Stem Cell Differentiation

Background:Extracellular vesicles (EVs) are derived from internal cellular compartments, and have potential as a diagnostic and therapeutic tool in degenerative disease associated with aging. Mesenchymal stem cells (MSCs) have become a promising tool for functional EVs production. This study investigated the efficacy of EVs and its effect on differentiation capacity.Methods:The characteristics of MSCs were evaluated by flow cytometry and stem cell differentiation analysis, and a production mode of functional EVs was scaled from MSCs. The concentration and size of EVs were quantitated by Nanoparticle Tracking Analysis (NTA). Western blot analysis was used to assess the protein expression of exosome-specific markers. The effects of MSC-derived EVs were assessed by chondrogenic and adipogenic differentiation analyses and histological observation.Results:The range of the particle size of adipose-derived stem cells (ADSCs)- and Wharton’s jelly -MSCs-derived EVs were from 130 to 150 nm as measured by NTA, which showed positive expression of exosomal markers. The chondrogenic induction ability was weakened in the absence of EVs in vitro. Interestingly, after EV administration, type II collagen, a major component in the cartilage extracellular matrix, was upregulated compared to the EV-free condition. Moreover, EVs decreased the lipid accumulation rate during adipogenic induction.Conclusion:The results indicated that the production model could facilitate production of effective EVs and further demonstrated the role of MSC-derived EVs in cell differentiation. MSC-derived EVs could be successfully used in cell-free therapy to guide chondrogenic differentiation of ADSC for future clinical applications in cartilage regeneration.

Cellular and Biochemical Characterization of Mesenchymal Stem Cells from Killian Nasal Polyp.

Killian's (antrochoanal) polyp is a unilateral nasal polypoid lesion of the maxillary sinus especially affecting children and young adults with unilateral nasal obstruction, pus discharge, and headache. Although its etiology is unclear, chronic inflammation, autoreactivity, allergies, and viral infections are implicated in its formation and development, causing nasal tissue remodeling. In this context, we isolated and cultured mesenchymal stem cells from surgical biopsies of three patients with Killian nasal polyp (KNP-MSCs) while healthy nasal tissue (HNT-MSCs) was used as control. Our results demonstrated that KNP-MSCs exhibited reduced cell proliferation compared to HNT-MSCs, and migrated less than the control, showing a partial epithelial phenotype with low mRNA levels of I-CAM and a significant increase of E-cad. Subsequently, both MSCs were induced to osteoblastic or adipocyte differentiation for up to 20 days. KNP-MSCs underwent to differentiate into osteoblasts but exhibited reduced ALP activity and calcium deposits and low mRNA levels of osteogenesis-associated genes compared to osteogenic induced-HNT-MSCs. Conversely, KNP-MSCs and HNT-MSCs have shown the same adipogenic differentiation potential, with a similar lipid droplet amount, adipocyte gene expression, and triacylglycerols content. Taken together, these results first demonstrated the cellular and molecular characterization of MSCs derived from the Killian nasal polyp.

Characterization of mesenchymal stem cells with augmented internalization of magnetic nanoparticles: The implication of therapeutic potential

Mesenchymal stem cells (MSCs) possess the potential to repair damaged tissue and regenerate the cells with a specific function. MSCs dispersed in the circulatory system tend to enter pulmonary tissue, arming the ability to change the lung microenvironment, prevent pulmonary fibrosis, and treat pulmonary injuries. It has been demonstrated that magnetic nanoparticles (MNPs) not only can be used as the vector for drug delivery, which can improve the specificity and control the drug released; but also act as the contract media for imaging, possessing the potential of being tracer and conducting differentiation to comply repairment. Epigallocatechin gallate (EGCG) has been known to greatly enhance MNP uptake by tumor cells under the influence of magnetic fields. However, whether EGCG significantly improves the MNP uptake by MSCs in the magnetic fields and whether MSCs maintain the characteristics of MSCs with such treatment remains unknown. In this study, we demonstrated that EGCG significantly enhanced the MNP uptake by MSCs in the magnetic field. With 30 µM EGCG in the magnetic field for 2 h, the cell-associated MNPs are increased 7.1 and 3.4 times by mouse and human MSCs without a distinguished influence on cell morphology, respectively. Furthermore, we evaluated the osteogenic differentiation potential of human MSCs after MNP uptake by assessing the relative gene expressions of osteogenic markers Runt-related transcription factor 2 (RUNX2), Osterix (Sp7), and Alkaline phosphatase (ALPL). There is no definite evidence showing that the expressions of these osteogenic marker genes, upon MNPs uptake, are significantly different from those of the control group. Surface antigen expression assay by flow cytometry revealed a similar expression population of human MSC surface markers CD73 and CD90. The results suggest that the stemness and differentiation capacity of MSCs are not affected after the MNP uptake. Our study demonstrates that combining EGCG and magnetic fields is an effective method to enhance MNP uptake by MSCs. The study serves as the foundation of the research related to the clinical application of combining MNPs and MSCs. We anticipate that the therapeutic potential of MSCs can be explored further to treat new coronary pneumonia and other diseases through this research platform.