Breast cancer remains a prevalent global health challenge, with tumor-removal surgeries being among the most common treatments but often leading to aesthetic defects. Adipose-derived stem cell (ADSC)-enriched fat grafting in breast reconstruction offers promising therapeutic benefits. However, concerns about its oncological safety persist, particularly regarding the potential risks of promoting cancer recurrence. This study investigated the effects of ADSCs on breast cancer progression by coculturing ADSCs with the MCF-7 breast cancer cell line for a short cell cultivation period of 3 days. We performed an RNA-seq analysis to identify significant transcriptomic changes in cocultured MCF-7 cells and carried out functional enrichment analyses to uncover key biological pathways influenced by ADSCs. Our findings revealed that transcriptomic alterations in MCF-7 cells are linked to aggressive cancer traits, including the upregulation of epithelial–mesenchymal transition (EMT) and the HIF-1 signaling pathway, which indicate a shift toward aerobic glycolysis. Some of the observed gene expression changes also correlated with relapse risk and mortality. These findings underscore the need for further research to explore the implications of these genes and pathways in driving aggressive cancer phenotypes and assess the safety of ADSCs in clinical settings.

Objectives: This study aimed to investigate the anti-inflammatory properties and underlying mechanisms of ostruthin, a natural coumarin isolated from the roots of Paramignya trimera . Using an in vitro model of co-stimulation with bacterial and viral mimetics in RAW264.7 macrophages, we examined the effects of ostruthin on the expression of key signaling molecules: Myd88 and Nf-κb1 (mediators of Toll-like receptor signaling), and Arid5a (a stabilizer of IL-6 mRNA). We also assessed the production of inflammatory cytokines, including IL-6, TNF-α, and IL-10. Methods: Ostruthin was isolated from P. trimera roots collected in Khanh Hoa Province, Vietnam. RAW264.7 cells were pre-treated with various concentrations of ostruthin before being stimulated with lipopolysaccharide (LPS) and poly(I:C). The mRNA expression of Myd88 , Nf-κb1 , and Arid5a was quantified at 3, 6, and 24 h using qRT-PCR. Cytokine levels were quantified at 24 h using ELISA. Additionally, molecular docking and molecular dynamics simulations were performed to predict direct interactions between ostruthin and the inflammation-related target, IL-6. Results: Ostruthin significantly and dose-dependently reduced IL-6, TNF-α, and IL-10 secretion in co-stimulated RAW264.7 cells, with IC ₅₀ values of 3.0 ± 1.2 µM, 4.8 ± 1.5 µM, and 11.3 ± 0.9 µM, respectively. Treatment with ostruthin (3-60 μM) also markedly downregulated Myd88 (by 36%-79%), Nf-κb1 (by 49%-97%), and Arid5a (by 29%-83%) mRNA expression relative to untreated controls. Docking simulations revealed favorable binding affinities of ostruthin with IL-6 (−5.381 kcal/mol), comparable to or exceeding those of known inhibitors like tocilizumab. Conclusions: Ostruthin exhibits potent anti-inflammatory activity in RAW264.7 macrophages stimulated with bacterial and viral mimetics, primarily by suppressing pro-inflammatory cytokines and modulating both TLR signaling and IL-6 mRNA stability pathways. These findings suggest that ostruthin may serve as a promising lead compound for the development of anti-inflammatory therapies targeting infections of mixed microbial origin.

Breast cancer is the most frequently diagnosed cancer in women globally. The tumor microenvironment plays a vital role in epithelial-to-mesenchymal transition (EMT), leading to the invasion and metastasis of cancer cells. The tumor microenvironment includes all components of the tumor cells, including the extracellular matrix, tumor vasculature, mesenchymal stem cells, immune cells, and fibroblasts. Understanding the interactions between mesenchymal stem cells and cancer cells is essential in determining the role of mesenchymal stem cells in diagnosing and treating breast cancer. In this study, we present the result of co-culture between adipose tissue mesenchymal stem cells (ADMSCs) and breast cancer cells (MCF-7 cell line) and determine the expression levels of transcription factors involved in EMT, including Twist and Snail. The results showed that the proliferation of MCF-7 co-cultured with ADMSCs was not increased compared to MCF-7 mono-cultured. Determination of gene expression levels by qRT-PCR revealed a significant increase in the EMT-related transcription factors (Twist and Snail) in breast cancer cells upon co-culture with ADMSCs. There were also significant differences between the expression levels of IL-6 and AhR in MCF-7 cells co-cultured with ADMSCs and MCF-7 cells mono-cultured. The results suggested that ADMSCs promoted the EMT of MCF-7 cells, potentially via AhR/NF-κB pathways.

Umbilical cord hematopoietic stem cells (UC-HSCs) are capable of self-regeneration and differentiation into many different types of blood cells, helping to treat blood diseases such as leukemia, Hodgkin's disease and non-Hodgkin's lymphoma, as well as for transplantation. However, the number of UC-HSCs is still very small due to limitations in isolation and proliferation. In this study, UC-HSCs were cultured with the bioactive SL-13R peptide to examine whether SL-13R enhances UC-HSC proliferation. Here, we showed that CD34+UC-HSCs can be enriched by up to 47.08% by culturing total mononuclear cells (MNCs) from umbilical cord blood (UCB) in serum-free StemMACS medium supplemented with a cytokine cocktail. We demonstrated that SL-13R enhanced the proliferation of CD34+UC-HSCs by 1.21-1.44 times compared to that of the controls. These results suggested that StemMACS medium supplemented with cytokines and SL-13R can be used to expand CD34+UC-HSCs ex vivo for medical treatment and transplantation.

Mesenchymal stem cells (MSCs) have garnered significant attention in biomedical studies due to their remarkable properties, such as self-renewal, differentiation into diverse cell types and immune responses. The proliferation and differentiation of MSCs are significantly influenced by the ligand-dependent transcription factor known as the aryl hydrocarbon receptor (AhR). In order to understand the roles of Ahr in adipose-derived MSCs (AD-MSCs), we disrupted the Ahr gene in this work using CRISPR/Cas9 gene editing technology. The gRNA/Cas9 dual vector and donor vector were introduced into the AD-MSC cell line (PT-5006). Green fluorescent protein (GFP) expression and puromycin resistance were used to identify the transfected cells. AhR-KO cells were cloned and confirmed by PCR and sequencing. By using RT-qPCR, the expression levels of AhR and the Ahr-related gene Cyp1B1 were investigated. The results showed that the knocked out of AhR using CRISPR/Cas9, resulting significantly decreased expression of 7.69-fold for AhR and 3.70-fold for Cyp1B1 in the cells. These cell clones and CRISPR/Cas9 vectors could be used as tools to investigate the functions of AhR in both AD-MSCs and other cell types.

There are three groups of potential stem cells that are widely used in regenerative medicine in clinical practice: hematopoietic stem cells, mesenchymal stem cells, and epithelial stem cells. Among them, adipose-derived mesenchymal stem cells (ASCs) are very common in the human body and can be easily obtained without causing major damage to the body. The use of human adipose-derived stromal vascular fraction (SVF) therapy has increased in recent years. SVF is a rich source of mesenchymal stem cells (MSCs) used in regenerative medicine. This study evaluated the effect of collagenase incubation time on the efficiency of SVF isolation and MSC viability. The aim of the study was to optimize the SVF isolation time to obtain MSCs with a high survival rate in clinical applications while ensuring the stemness and differentiation ability of MSCs after isolation. Collagenase digestion at different time points (60, 120, and 180 minutes) was compared, measuring MSC yield and viability via flow cytometry and differentiation assays. Results indicated that 120 minutes of collagenase treatment provided optimal MSC yield and cell viability, demonstrating significant differentiation potential, making this time point ideal for clinical applications.