• Adipose-derived stem cells
• heterogeneity
• immunophenotypes
• personalized medicine
• subpopulation selection

• biodegradables
• self-reinforcing
• small diameter vascular grafts
• tissue engineering

• Rats
• Animals
• Tissue Engineering
• X-Ray Microtomography
• Blood Vessel Prosthesis
• Vascular Grafting
• Polyurethanes

Multiple studies have demonstrated that various nanoparticles (NPs) stimulate osteogenic differentiation of mesenchymal stem cells (MSCs) and inhibit adipogenic ones. The mechanisms of these effects are not determined. The aim of this paper was to estimate Wharton’s Jelly MSCs phenotype and humoral factor production during tri-lineage differentiation per se and in the presence of silicon–gold NPs. Silicon (SiNPs), gold (AuNPs), and 10% Au-doped Si nanoparticles (SiAuNPs) were synthesized by laser ablation, characterized, and studied in MSC cultures before and during differentiation. Humoral factor production (n = 41) was analyzed by Luminex technology. NPs were nontoxic, did not induce ROS production, and stimulated G-CSF, GM-CSF, VEGF, CXCL1 (GRO) production in four day MSC cultures. During MSC differentiation, all NPs stimulated CD13 and CD90 expression in osteogenic cultures. MSC differentiation resulted in a decrease in multiple humoral factor production to day 14 of incubation. NPs did not significantly affect the production in chondrogenic cultures and stimulated it in both osteogenic and adipogenic ones. The major difference in the protein production between osteogenic and adipogenic MSC cultures in the presence of NPs was VEGF level, which was unaffected in osteogenic cells and 4–9 times increased in adipogenic ones. The effects of NPs decreased in a row AuNPs > SiAuNPs > SiNPs. Taken collectively, high expression of CD13 and CD90 by MSCs and critical level of VEGF production can, at least, partially explain the stimulatory effect of NPs on MSC osteogenic differentiation.

• G-CSG
• Luminex
• MSC
• VEGF
• Wharton’s jelly mesenchymal stem cells
• adipogenic differentiation
• cell phenotype
• chondrogenic
• osteogenic
• silicon–gold nanoparticles
• soluble factors

• partially deproteinized biologic bone
• fibronectin
• vascular endothelial cells
• adipose‑derived stem cells
• co‑culture system

• Adipose Tissue/cytology
• Adipose Tissue/growth & development
• Adipose Tissue/metabolism
• Animals
• Bone and Bones/cytology
• Bone and Bones/metabolism
• Cell Adhesion
• Cell Differentiation
• Cell Survival
• Cells, Cultured
• Coculture Techniques/methods
• Endothelial Cells/cytology
• Endothelial Cells/metabolism
• Female
• Fetal Blood
• Fibronectins/metabolism
• Fluorescent Antibody Technique
• Microscopy, Electron, Scanning
• Rats, Sprague-Dawley
• Stem Cells/cytology
• Stem Cells/metabolism
• Time Factors
• Tissue Engineering/methods
• Tissue Scaffolds
• Rats

Organ transplantation is a standard therapeutic strategy for irreversible organ damage, but the utility of nerve transplantation remains generally unexplored, despite its potential benefit to a large patient population. Here, we aimed to establish a feasible preclinical mouse model for understanding the cellular mechanisms behind the rejection of peripheral and optic nerves.

We performed syngenic and allogenic transplantation of optic and sciatic nerves in mice by inserting the nerve grafts inside the kidney capsule, and we assessed the allografts for signs of rejection through 14 d following transplantation. Then, we assessed the efficacy of CTLA4 Ig, Rapamycin, and anti-CD3 antibody in suppressing immune cell infiltration of the nerve allografts.

By 3 d posttransplantation, both sciatic and optic nerves transplanted from BALB/c mice into C57BL/6J recipients contained immune cell infiltrates, which included more CD11b⁺ macrophages than CD3⁺ T cells or B220⁺ B cells. Ex vivo immunogenicity assays demonstrated that sciatic nerves demonstrated higher alloreactivity in comparison with optic nerves. Interestingly, optic nerves contained higher populations of anti-inflammatory PD-L1⁺ cells than sciatic nerves. Treatment with anti-CD3 antibody reduced immune cell infiltrates in the optic nerve allograft, but exerted no significant effect in the sciatic nerve allograft.

These findings establish the feasibility of a preclinical allogenic nerve transplantation model and provide the basis for future testing of directed, high-intensity immunosuppression in these mice.

Adipose-derived stem cells (ADSCs) are promising cell sources for regenerative medicine due to the simplicity of their harvest and culture; however, their biological properties are not completely understood. Moreover, recent murine and human studies identified several functional subpopulations of ADSCs varying in differentiation potential; however, there is a lack of research on canine ADSCs. Cystine transporter (xCT) is a stem cell marker in gastric and colon cancers that interacts with CD44 to enhance cystine uptake from the cell surface and subsequently accelerates intercellular glutathione levels. In this study, we identified a ~5% functional subpopulation of canine ADSCs with xCT⁺ expression (xCT^Hi). Compared with those of the xCT⁻ subpopulation (xCT^Lo), the xCT^Hi subpopulation showed a significantly higher proliferation rate, higher expression of conventional stem cell markers (SOX2, KLF4, and c-Myc), and higher expression of adipogenic markers (FABP4 and PPARγ). By contrast, the xCT^Lo subpopulation showed significantly higher expression of osteogenic markers (BMP1 and SPP) than xCT^Hi cells. These results suggest xCT as a candidate marker for detecting a functional subpopulation of canine ADSCs. Mechanistically, xCT could increase the adipogenic potential while decreasing the osteogenic differentiation potential, which could serve as a valuable target marker in regenerative veterinary medicine.

• adipogenic differentiation
• adipose-derived stem cell
• cystine transporter
• flow cytometry
• osteogenic differentiation

• conditioned medium
• epigenetic reprogramming
• pancreatic β cell

To identify and characterize functionally distinct subpopulation of adipose-derived stem cells (ADSCs).

ADSCs cultured from mouse subcutaneous adipose tissue were sorted fluorescence-activated cell sorter based on aldehyde dehydrogenase (ALDH) activity, a widely used stem cell marker. Differentiation potentials were analyzed by utilizing immunocytofluorescece and its quantitative analysis.

Approximately 15% of bulk ADSCs showed high ALDH activity in flow cytometric analysis. Although significant difference was not seen in proliferation capacity, the adipogenic and osteogenic differentiation capacity was higher in ALDH^Hi subpopulations than in ALDH^Lo. Gene set enrichment analysis revealed that ribosome-related gene sets were enriched in the ALDH^Hi subpopulation.

High ALDH activity is a useful marker for identifying functionally different subpopulations in murine ADSCs. Additionally, we suggested the importance of ribosome for differentiation of ADSCs by gene set enrichment analysis.

• Adipose-derived stem/stromal cell
• Aldehyde dehydrogenase activity
• Flow cytometry
• Subpopulation
• Ribosome

• Cartilage
• Chondrogenesis
• Dermal fibroblast
• Extracellular matrix
• Mesenchymal stem cell

Adipose-derived stem cells (ADSCs) are abundant and readily obtained, and have been studied for their clinical applicability in regenerative medicine. Some surface antigens have been identified as markers of different ADSC subpopulations in mice and humans. However, it is unclear whether functionally distinct subpopulations exist in dogs. To address this issue, we evaluated aldehyde dehydrogenase (ALDH) activity—a widely used stem cell marker in mice and humans—by flow cytometry. Approximately 20% of bulk ADSCs showed high ALDH activity. Compared to cells with low activity (ALDH^Lo), the high-activity (ALDH^Hi) subpopulation exhibited a higher capacity for adipogenic and osteogenic differentiation. This is the first report of distinct ADSC subpopulations in dogs that differ in terms of adipogenic and osteogenic differentiation potential.

• adipogenic differentiation
• adipose-derived stem cells
• aldehyde dehydrogenase activity
• flow cytometry
• osteogenic differentiation

• CD90(+)
• New bone formation
• Periosteum
• Periosteum-derived cell
• Three-dimensional cell migration

• Animals
• Biomarkers/metabolism
• Bone Marrow Cells/cytology
• Bone Marrow Cells/drug effects
• Bone Marrow Cells/metabolism
• Bone Regeneration/physiology
• Cell Differentiation/drug effects
• Cell Movement/drug effects
• Cell Proliferation/drug effects
• Collagen/chemistry
• Culture Media/chemistry
• Culture Media/pharmacology
• Female
• Femur/injuries
• Flow Cytometry
• Gelatin/chemistry
• Mesenchymal Stem Cell Transplantation
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cells/drug effects
• Mesenchymal Stem Cells/metabolism
• Mice
• Mice, Inbred ICR
• Osteogenesis/physiology
• Periosteum/cytology
• Periosteum/drug effects
• Periosteum/metabolism
• Primary Cell Culture
• Thy-1 Antigens/metabolism
• Tissue Engineering
• Tissue Scaffolds

Adipose tissue is a rich, ubiquitous and easily accessible source for multipotent stromal/stem cells and has, therefore, several advantages compared to other sources of mesenchymal stromal/stem cells. Several studies have tried to identify the origin of the stromal/stem cell population within adipose tissue in situ. This is a complicated attempt because no marker has currently been described which unambiguously identifies native adipose-derived stromal/stem cells (ASCs). Isolated and cultured ASCs are a non-uniform preparation consisting of several subsets of stem and precursor cells. Cultured ASCs are characterized by their expression of a panel of markers (and the absence of others), whereas their in vitro phenotype is dynamic. Some markers were expressed de novo during culture, the expression of some markers is lost. For a long time, CD34 expression was solely used to characterize haematopoietic stem and progenitor cells, but now it has become evident that it is also a potential marker to identify an ASC subpopulation in situ and after a short culture time. Nevertheless, long-term cultured ASCs do not express CD34, perhaps due to the artificial environment. This review gives an update of the recently published data on the origin and phenotype of ASCs both in vivo and in vitro. In addition, the composition of ASCs (or their subpopulations) seems to vary between different laboratories and preparations. This heterogeneity of ASC preparations may result from different reasons. One of the main problems in comparing results from different laboratories is the lack of a standardized isolation and culture protocol for ASCs. Since many aspects of ASCs, such as the differential potential or the current use in clinical trials, are fully described in other recent reviews, this review further updates the more basic research issues concerning ASCs’ subpopulations, heterogeneity and culture standardization.

• Adipose-derived stromal/stem cells
• Adipose tissue
• Subpopulation
• Heterogeneity
• Phenotype
• CD34
• Mesenchymal stem cells

• Bone marrow
• emphysema
• mesenchymal stem cells
• transplantation

• Mesenchymal stem cells
• repair
• spinal nerves
• transplantation