Adipocyte-derived Stem Cells Research Articles

Extracellular vesicle transfer of miR-1 to adipose tissue modifies lipolytic pathways following resistance exercise.

Extracellular vesicles (EVs) have emerged as important mediators of intertissue signaling and exercise adaptations. In this human study, we provide evidence that muscle-specific microRNA-1 (miR-1) was transferred to adipose tissue via EVs following an acute bout of resistance exercise. Using a multimodel machine learning automation tool, we discovered muscle primary miR-1 transcript and CD63+ EV count in circulation as top explanatory features for changes in adipose miR-1 levels in response to resistance exercise. RNA-Seq and in-silico prediction of miR-1 target genes identified caveolin 2 (CAV2) and tripartite motif containing 6 (TRIM6) as miR-1 target genes downregulated in the adipose tissue of a subset of participants with the highest increases in miR-1 levels following resistance exercise. Overexpression of miR-1 in differentiated human adipocyte-derived stem cells downregulated these miR-1 targets and enhanced catecholamine-induced lipolysis. These data identify a potential EV-mediated mechanism by which skeletal muscle communicates with adipose tissue and modulates lipolysis via miR-1.

Enrichment of Fat Graft with Association of ASC and Nanofat in an Animal Model.

Fat graft (FG) absorption rate varies from 20 to 80% in two years. Recently, several bioengineering techniques were applied to improve FG retention rate. Numerous studies investigated the use of adipocyte-derived stem cells (ASC) as FG enrichment. However, ASC production is costly, complex, and time-consuming. In contrast, Nanofat, a combination of lipids, stem cells and growth factors, offers a faster, simpler, and more cost-effective alternative for FG enrichment. This study aims to compare the effects of ASC with those of Nanofat, as a viable option in FG enrichment. Animals were allocated in three groups: Control group (1mL fat), ASC group (1mL fat +1x106ASC), andNnF group (1mL of fat + 0.3mL NnF). These groups were subdivided in three subgroups (4, 8, and 12 weeks, n = 6/group). We performed ultrasound and macroscopic measurements for FG volume, histology and expression of healing and inflammation genes. At week 12, ASC and NnF groups showed a higher retention of FG when compared to the Control group (51%, 46%, 12% respectively, p < 0.01). Fibrosis was similar in ASC and Nanofat groups. The Nanofat group showed a higher vascular density then the Control group (p < 0.05). Il-10 gene expression was higher, and Mmp9 was lower in the Nanofat group when compared to the ASC and Control groups. This study indicates that enriching FG with both ASC and Nanofat led to an increased retention rate of the FG, suggesting that Nanofat might be a promising alternative for FG enrichment. This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

220 Strontium enhances in vitro osteogenic differentiation of porcine adipocyte–derived stem cells

220 Strontium enhances in vitro osteogenic differentiation of porcine adipocyte–derived stem cells

Lipofilling-A Regenerative Alternate for Remodeling Burn Scars: A Clinico-Immunohistopathological Study.

Introduction Any injury involving the dermis will lead to scarring. Scar tissue can cause functional limitations, cosmetic impairments, pain, and itch. Adipose-derived stem cells have also been shown to play a role in scar modulation. This study evaluates changes in lipofilled scar over the period of time and compares it with non-lipofilled scar tissue. Materials and Methods A prospective case-control study with intraindividual follow-up was performed on 30 adult patients with post-burn scars from November 2016 to May 2019. Clinical, histopathological, and immunohistochemical parameters were assessed among the case and control regions of the scar. Results Mean age of the study population was 30.6 years. The duration of the scar included in this study ranged from 1 to 28 years, with a mean duration of 5.91 years. There was a significant reduction in pain, itch, stiffness, and an increase in the pliability of the scar, and a substantial improvement in the modified Vancouver Scar Score in the lipofilled group. In histopathological analysis, the case group showed organized parallel collagen fibers, a significant reduction in melanocytes, improvement in vascularity, and a significantly increased amount of collagen fibers at the reticular dermis. Immunohistochemical analysis indicated new cell synthesis in the scar tissue and reduced melanocytes. Conclusion The remodeling effect of adipocyte-derived stem cells is long-lasting, and there is a gradual improvement in most of the parameters. Lipofilling has regenerative capacity, which leads to the improved overall appearance of scar and improvement at the cellular level.

125. Effect of Hyperbaric Oxygen Chamber Associated with the Use of Adipocyte-derived Stem Cells in the Treatment of Degloving Injuries: Experimental Study in Rats

125. Effect of Hyperbaric Oxygen Chamber Associated with the Use of Adipocyte-derived Stem Cells in the Treatment of Degloving Injuries: Experimental Study in Rats

Abstract 1464: Exosomal delivery of tumor suppressive microRNA-7 as a novel approach to regulating cervical cancer cell growth

Abstract There is an unmet need for more safe, effective, targeted therapy in cervical cancer which this study seeks to meet through the use of exosomes and miRNA. Although cervical cancer is the fourth most prevalent female cancer worldwide, it currently has sparce options for treatment. Early-stage tumors are often surgically removed, while later stage cancers are treated with traditional radiation and chemotherapy methods, which have limited efficacy. Thus, to bridge this gap, we propose the use of exosomes, which are naturally occurring cell-derived vesicles, in combination with microRNA-7 (miR-7), a known tumor-suppressive microRNA, as a potential treatment for cervical cancer. We first transfected adipocyte-derived stem cells (ADSC) with miR-7. ADSC produce exosomes that are immune-compatible, have a long half-life, and have an especially low toxicity, making them a versatile nanocarrier. We collected the conditioned media produced by the ADSC, from which we isolated the miR-7-containing exosomes. The collected exosomes were confirmed to contain the appropriate miRNA by qPCR and applied to HeLa cells, and cellular response and gene expression changes were analyzed via qPCR. Functional assays including proliferation and migration were also performed to analyze the effect of exosome treatment on cell function. Controls for this study included a parallel ADSC transfection with a non-coding, scramble miRNA, as well as HeLa cells treated with exosomes from non-transfected ADSC. Our studies demonstrate that transfection of ADSC with miR-7 produces exosomes with significantly increased levels of this regulatory miRNA, and that these exosomes can specifically deliver their cargo and effectively downregulate the target gene Epidermal Growth Factor Receptor (EGFR), which is involved in tumor progression in cervical cancer. Additionally, we show that these miR-7-containing exosomes decrease proliferation and migration of HeLa cells in our in vitro functional analysis. Ultimately, this technology provides an alternative treatment option that will improve clinical outcomes of women who are affected by cervical cancer. Citation Format: Gabriella McWilliams, Molly Roy, Daniel Bilbao Cortes, Yu-Ping Yang, Sylvia Daunert, Sapna Deo. Exosomal delivery of tumor suppressive microRNA-7 as a novel approach to regulating cervical cancer cell growth [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1464.

Therapeutic Effect of Human Adipocyte-derived Stem Cell-derived Exosomes on a Transgenic Mouse Model of Parkinson's Disease.

Stem cell therapy and regenerative medicine are promising for treating Parkinson's disease (PD) not only for the potential for cell replacement but also for the paracrine effect of stem cell secretion, especially proteins and nucleotide-enriched exosomes. This study investigated the neuroprotective effect of exosomes secreted from human adipocyte-derived stem cells (hADSCs) on PD. hADSCs were isolated from the visceral fat tissue of individuals without PD who underwent bariatric surgery and were validated using surface markers and differentiation ability. Exosomes were isolated from the culture medium of hADSCs through serial ultracentrifugation and validated. Condensed exosomes were administered intravenously to 12-week-old MitoPark mice, transgenic parkinsonism mouse model with conditional knockout of mitochondrial transcription factor A in dopaminergic neurons, monthly for 3 months. Motor function, gait, and memory were assessed monthly, and immunohistochemical analysis of neuronal and inflammatory markers was performed at the end of the experiments. The hADSC-derived exosome-treated mice exhibited better motor function in beam walking and gait analyses than did the untreated mice. In the novel object recognition tests, the exosome-treated mice retained better memory function. Immunohistochemical analysis revealed that although exosome treatment did not prevent the loss of dopaminergic neurons in the substantia nigra of mice, it down-regulated microglial activation and neuroinflammation in the midbrain. hADSC-derived exosomes were neuroprotective in this in vivo mouse model of PD, likely because of their anti-inflammatory effect. Use of hADSC-derived exosomes may offer several beneficial effects in stem cell therapy. Since they can also be produced at an industrial level, they are a promising treatment option for PD and other neurodegenerative diseases.

Human T2D-Associated Gene IMP2/IGF2BP2 Promotes the Commitment of Mesenchymal Stem Cells Into Adipogenic Lineage.

Excessive adiposity is the main cause of obesity and type two diabetes (T2D). Variants in human IMP2/IGF2BP2 gene are associated with increased risk of T2D. However, little is known about its role in adipogenesis and in insulin resistance. Here, we investigate the function of IMP2 during adipocyte development. Mice with Imp2 deletion in mesenchymal stem cells (MSC) are resistant to diet-induced obesity without glucose and insulin tolerance affected. Imp2 is essential for the early commitment of adipocyte-derived stem cells (ADSC) into preadipocytes, but the deletion of Imp2 in MSC is not required for the proliferation and terminal differentiation of committed preadipocytes. Mechanistically, Imp2 binds Wnt receptor Fzd8 mRNA and promotes its degradation by recruiting CCR4-NOT deadenylase complex in an mTOR-dependent manner. Our data demonstrate that Imp2 is required for maintaining white adipose tissue homeostasis through controlling mRNA stability in ADSC. However, the contribution of IMP2 to insulin resistance, a main risk of T2D, is not evident.

Ceiling culture of human mature white adipocytes with a browning agent: A novel approach to induce transdifferentiation into beige adipocytes.

Excess and dysfunctional adipose tissue plays an important role in metabolic diseases, including obesity, atherosclerosis and type 2 diabetes mellitus. In mammals, adipose tissue is categorized into two types: white and brown. Adult brown tissue is mainly composed of beige adipocytes, which dispose of stored energy as heat and have become increasingly popular as a therapeutic target for obesity. However, there is still a paucity of cell models that allow transdifferentiation of mature white adipocytes into beige adipocytes, as seen in vivo. Here, we describe a novel, ceiling culture-based model of human mature white adipocytes, which transdifferentiate into beige adipocytes under the mechanical force and hypoxia of ceiling culture. We also show that the use of rosiglitazone and rapamycin can modulate transdifferentiation, up and down regulating expression of beige adipocyte-specific genes, respectively. Rosiglitazone additionally facilitated the upregulation of fatty acid lipolysis and oxidation genes. Finally, these beige adipocytes derived from dedifferentiated adipocytes exhibited a progenitor-specific phenotype, with higher expression of mature adipocyte-specific genes than adipocyte-derived stem cells. Overall, we report a novel approach to conveniently cultivate beige adipocytes from white adipocytes in vitro, suitable for mechanistic studies of adipose biology and development of cell and drug therapies in the future.

Exosome delivery of tumor suppressive microRNA-7 to cervical cancer cells: A novel approach for regulating cervical cancer cell growth (230)

<b>Objectives:</b> Exosomes are highly stable nanosized extracellular vehicles released from cells for cell-cell communication that protect their cargo from degradation in an array of environments. Adipocytederived stem cell exosomes (ACS-exos) have unique properties that allow them to be engineered as delivery vehicles in various biological settings. The ability to manipulate the exosomes and their contents allows exosomes to be a focus of cancer therapeutics. MicroRNAs (miRNAs) are small non-coding RNAs that control gene translation and expression. Abnormal expression of tissue miRNAs has been associated with significant alterations in cell growth. MicroRNA-7 (miR-7) has an inhibitory effect on X-Linked Inhibitor of Apoptosis Protein (XIAP), which is over-expressed in cervical cancer cells, inhibiting apoptosis and promoting cell proliferation. miR-7 down-regulates XIAP in cervical carcinoma cells, increasing apoptosis and suppressing colony formation. Our objective was to develop a new therapeutic approach for cervical cancer by employing ACS-exos to deliver miR-7 to cervical cancer cells so as to elicit regulatory effects on XIAP. <b>Methods:</b> The cervical cancer cell line C-33 A was used to demonstrate the effect of miR-7 on XIAP expression. An miRNA mimic hsa-miR-7 was used for transfection. The presence of XIAP in C-33 A cells was established using Western blot, and then the cells were transfected with miR-7 using Lipofectamine ^TM RNAiMAX. The transfected cells were collected 24 and 48 hours after treatment, and the expression of XIAP was tested using Western blot. To collect exosomes, adipocyte-derived stem cells (ACS) were cultured in a growth medium with exosome-depleted FBS for 72 hours, followed by differential centrifugation. The isolated exosomes were then transfected with either miR-7 or Texas-Red fluorescent-labeled siRNA (TX-Red) using Exo-Fect^TM Exosome Transfection Reagent. Transfected exosomes were added to the C-33 A cell culture media. The cells were analyzed under a fluorescence microscope after 24 and 48 hours of growth. Western blot was used to study the expression of XIAP. The presence of Beta-Tubulin was also tested as a loading control. <b>Results:</b> The presence of XIAP in C-33 A cells was confirmed by Western blot. In the cells transfected with miR-7, Western blot demonstrated downregulation of XIAP after both 24 and 48 hours. Successful nucleotide transfection of ACS-exos and oligonucleotide integration into the C-33 A cells was confirmed by fluorescence microscopy, which demonstrated the intracellular presence of TX-Red in only the positive control C-33 A cell line. A similar downregulation of XIAP in the cells treated with miR-7 transfected ACS-exos was seen as in the miR-7 transfected cells by Western blot. Beta-Tubulin expression was seen across the control and treatment cells. <b>Conclusions:</b> Delivery of tumor-suppressive miR-7 via ACS-exos elicited downregulation of XIAP. The isolation and successful transfection of exosomes demonstrated are key processes in utilizing exosomes for cancer therapeutics. Further, the exosome cargo nucleotides integrated into the target cells and resulted in alteration of protein expression. Additional studies examining the effects of exosome-delivered miR-7 on cervical cancer cell viability, proliferation, invasion are underway. These findings highlight the promising therapeutic potential of exosomes in cervical cancer.

Molecular changes in adipocyte-derived stem cells during their interplay with cervical cancer cells.

Obesity is as an important risk factor and has been associated with a worse prognosis in at least 13 distinct tumor types. This is partially due to intercellular communication between tumor cells and adipose tissue-derived stem cells (ADSCs), which are increased in obese individuals. As yet, however, little is known about the molecular changes occurring in ADSCs in these conditions. Cervical cancer has a high incidence and mortality rate in women from developing countries, particularly in those with a high body mass index (BMI). We analyzed the expression profile of ADSCs co-cultured with cervical cancer cells through massive RNA sequencing followed by evaluation of various functional alterations resulting from the modified transcriptome. A total of 761 coding and non-coding dysregulated RNAs were identified in ADSCs after co-culture with HeLa cells (validation in CaSki and SiHA cells). Subsequent network analysis showed that these changes were correlated with migration, stemness, DNA repair and cytokine production. Functional experiments revealed a larger ALDHhigh subpopulation and a higher migrative capacity of ADSCs after co-culture with HeLa cells. Interestingly, CXCL3 and its intragenic long-noncoding RNA, lnc-CXCL3, were found to be co-regulated during co-culture. A loss-of-function assay revealed that lnc-CXCL3 acts as a key regulator of CXCL3 expression. Our results suggest that intercellular communication between ADSCs and cervical cancer cells modifies the RNA expression profile in the former, including that of lncRNAs, which in turn can regulate the expression of diverse chemokines that favor malignancy-associated capacities such as migration.

The impact of obesity on adipocyte-derived extracellular vesicles.

Recently, the emerging roles of adipocyte-derived extracellular vesicles (EVs) linking obesity and its comorbidities have been recognized. In obese subjects, adipocytes are having hypertrophic growth and are under stressed. The dysfunction adipocytes dysregulate the assembly of the biological components in the EVs including exosomes. This article critically reviews the current findings on the impact of obesity on the exosomal cargo contents that induce the pathophysiological changes. Besides, this review also summarizes the understanding on how obesity affects the biogenesis of adipocyte-derived exosomes and the exosome secretion. Furthermore, the differences of the exosomal contents in different adipose depots, and the impact of obesity on the exosomes that are derived from the stromal vascular fraction such as the adipose tissue macrophages and adipocyte-derived stem cells will also be discussed. The current development and potential application of exosome-based therapy will be summarized. This review provides crucial information for the design of novel exosome-based therapy for the treatment of obesity and its comorbidities.

Intravenous SPION-labeled adipocyte-derived stem cells targeted to the brain by magnetic attraction in a rat stroke model: An ultrastructural insight into cell fate within the brain

Mesenchymal stem cell therapy after stroke is a promising option investigated in animal models and clinical trials. The intravenous route is commonly used in clinical settings guaranteeing an adequate safety profile although low yields of engraftment. In this report, rats subjected to ischemic stroke were injected with adipose-derived stem cells (ADSCs) labeled with superparamagnetic iron oxide nanoparticles (SPIONs) applying an external magnetic field in the skull to retain the cells. Although most published studies demonstrate viability of ADSCs, only a few have used ultrastructural techniques. In our study, the application of a local magnetic force resulted in a tendency for higher yields of SPION-ADSCs targeting the brain. However, grafted cells displayed morphological signs of death, one day after administration, and correlative microscopy showed active microglia and astrocytes associated in the process of scavenging. Thus, we conclude that, although successfully targeted within the brain, SPION-ADSCs viability was rapidly compromised.

Clinical efficacy of adipocyte-derived stem cells conditioned media combined with micro-injury in refractory patch of alopecia areata.

Management of alopecia areata (AA) is often challenging especially when patients have AA lesion refractory to conventional treatments such as corticosteroids, contact immunotherapy, and systemic therapy. Reports indicate adipocyte-derived stem cell conditioned media (ASC-CM) can activate hair growth and micro-injury using fractional laser or microneedling can also induce wound healing and hair regeneration, which suggests ASC-CM combined with fractional laser or microneedling might provide alternative therapeutic option for a refractory patch of AA. This study aimed to evaluate the clinical efficacy and safety of ASC-CM combined with 10,600nm carbon dioxide fractional laser or microneedling for the treatment of refractory patch of AA. This retrospective study was based on evaluations of 14 patients with a refractory patch of AA treated with ASC-CM, combined with a 10,600nm carbon dioxide fractional laser, or microneedling from March 2017 to August 2020. The efficacy of treatment was assessed by extents of hair regrowth percentages of involved areas. Of the 14 enrolled patients, 9 (64.3%) showed > 50% hair regrowth and 6 patients (42.9%) showed complete recovery. In the responder group (n = 9), mean period to achieve > 50% hair regrowth was 11.3weeks (range 8-16weeks). In the non-responder group (n = 5), 4 patients (28.6%) showed < 25% of hair regrowth and 1 patient show slight hair regrowth (7.1%) after 3months of treatment. This study showed ASC-CM combined with 10,600nm carbon dioxide fractional laser or microneedling may offer effective and safe treatment options for a refractory patch of AA.

Developing a 3D B Cell Lymphoma Culture System to Model Antibody Therapy.

Diffuse large cell B cell lymphoma (DLBCL) accounts for approximately 30%–40% of all non-Hodgkin lymphoma (NHL) cases. Current first line DLBCL treatment results in long-term remission in more than 60% of cases. However, those patients with primary refractory disease or early relapse exhibit poor prognosis, highlighting a requirement for alternative therapies. Our aim was to develop a novel model of DLBCL that facilitates in vitro testing of current and novel therapies by replicating key components of the tumor microenvironment (TME) in a three-dimensional (3D) culture system that would enable primary DLBCL cell survival and study ex vivo. The TME is a complex ecosystem, comprising malignant and non-malignant cells, including cancer-associated fibroblasts (CAF) and tumor-associated macrophages (TAM) whose reciprocal crosstalk drives tumor initiation and growth while fostering an immunosuppressive milieu enabling its persistence. The requirement to recapitulate, at least to some degree, this complex, interactive network is exemplified by the rapid cell death of primary DLBCL cells removed from their TME and cultured alone in vitro. Building on previously described methodologies to generate lymphoid-like fibroblasts from adipocyte derived stem cells (ADSC), we confirmed lymphocytes, specifically B cells, interacted with this ADSC-derived stroma, in the presence or absence of monocyte-derived macrophages (MDM), in both two-dimensional (2D) cultures and a 3D collagen-based spheroid system. Furthermore, we demonstrated that DLBCL cells cultured in this system interact with its constituent components, resulting in their improved viability as compared to ex-vivo 2D monocultures. We then assessed the utility of this system as a platform to study therapeutics in the context of antibody-directed phagocytosis, using rituximab as a model immunotherapeutic antibody. Overall, we describe a novel 3D spheroid co-culture system comprising key components of the DLBCL TME with the potential to serve as a testbed for novel therapeutics, targeting key cellular constituents of the TME, such as CAF and/or TAM.

The influence of adipocyte-derived stem cells (ASCs) on the ischemic epigastric flap survival in diabetic rats.

ABSTRACTPurpose:To assess the effects of adipocyte-derived stem cell (ASC)-injection on the survival of surgical flaps under ischemia in diabetic rats.Methods:Diabetes was induced in 30 male Wistar rats using streptozotocin (55 mg/kg). After eight weeks, epigastric flap (EF) surgery was performed. The animals were divided into control (CG), medium-solution (MG), and ASC groups. The outcomes were: the survival area (SA), the survival/total area rate (S/TR), and expression levels (EL) of genes: C5ar1, Icam1, Nos2, Vegf-a.Results:In the ASC group, compared to CG, we observed improved flap SA (CG-420 mm2 vs. ASC-720 mm2; p=0.003) was observed. The S/TR analysis was larger in the ASC group (78%) than the CG (45%). This study showed an increase in the Vegf-a EL in the ASC group (2.3) vs. CG (0.93, p=0.0008). The Nos2 EL increased four-fold in the ASC group compared to CG, and C5ar1 EL decreased almost two-fold in the ASC group vs. the CG (p=0.02). There was no difference among the groups regarding Icam1 EL. Compared to the MG, the ASC group had a bigger flap SA (720 mm2 vs. 301 mm2, respectively), a bigger S/TR (78% vs. 32%, p=0.06, respectively) and increased EL of Vegf-a (2.3 vs. 1.3, respectively). No difference between ASC-group and MG was seen regarding Nos2 (p=0.08) and C5ar1 (p=0.05).Conclusions:This study suggests that ASCs increase the survival of EF under IR in diabetic rats.

Protocol to Study the Role of Extracellular Vesicles During Induced Stem Cell Differentiation.

Extracellular vesicles (EVs) are vesicles released by cells, which due to their cargo and cell membrane proteins induce changes in the recipient cells. These vesicles can be a novel option to induce stem cell differentiation. Here we described a method to induce mesenchymal stem cell differentiation (MSC) into neuron-like cells using small EVs from neurons. First, we will describe a method based on neurons to induce adipocyte derived stem cells differentiation, a type of MSC, by coculturing both using inserts. Secondly, we will describe a follow-up method by using only isolated neuron-derived small EVs to directly induce ADSC differentiation in neuron-like cells. Importantly, in both methods it is possible to avoid the direct cell-to-cell contact, thus allowing for the study of soluble factors role during stem cell differentiation.

QPCR analysis of mesenchymal stem cell marker expression during the long-term culture of canine adipocyte derived stem cells

Abstract Due to its availability and accessibility, adipose tissue has been the subject of various studies in many different medical fields and is believed to be a useful source of stem cells. The ability of ASCs to differentiate towards different cell lineages, with possibility of directing this differentiation, increases their possible clinical applications, and they have been widely employed in multiple therapies and treatment of different pathologies. However, a deeper understanding of the molecular mechanisms underlying the ASCs osteoblastic and chondrocyte differentiation may lead to novel applications treating a multitude of different bone-related diseases through techniques more likely meeting worldwide consensus. In this study, the RT-qPCR method was used to determine the changes in expression of ASC specific markers (CD105, CD73, CD14, CD34, CD90 and CD45) before and after long-term (14-day) in vitro cultures. To confirm the identity of the investigated cells, flow cytometry was used to evaluate the presence of positive (CD44, CD90) and negative (CD45, CD34) ASC markers. Overall, the results of the PCR analysis showed a significant change in expression of most of the marker genes, indicating significant changes in the cultured cells caused by their long-term culture, potentially altering their original stem-like characteristics. Running title: ASC marker expression during long-term in vitro culture

Therapeutic Effect of Intravenously Administered Autologous Adipocyte-Derived Stem Cells on Chronic Stage Stroke Patients

Therapeutic Effect of Intravenously Administered Autologous Adipocyte-Derived Stem Cells on Chronic Stage Stroke Patients

Metastasis-on-a-chip reveals adipocyte-derived lipids trigger cancer cell migration via HIF-1α activation in cancer cells

Although obesity is a newly considered risk factor for cancer, the mechanisms by which adipocyte-derived metabolites accelerate cancer malignancy have yet to be elucidated. To identify the connection among heterogeneous cell types, conventional methods including Transwell assays or conditioned media (CM) have been used; however, these methods do not fully reflect niche effects in the tumor microenvironment (TME). Here, we established an oxygen permeable polydimethylsiloxane (PDMS)-based three-dimensional (3D) culture system to allow direct attachment between human adipocyte derived stem cells (ADSCs) and cancer cells. By doing so, a physiologically bioactive TME was created, which could be used to reveal further the relationships between different cell types. We found that co-culture of cancer cells with ADSCs resulted in a dispersion phenomenon, and the dispersed spheroid was well matched with the enhanced metastatic potential of cancer cells. Lipid profiling and in vitro migration assays suggested that lipids are the driving force for cancer cell migration via HIF-1α upregulation. In addition, the lipid/HIF-1α axis promoted tumor metastasis in a xenograft mouse model. This study presents an in vitro model of a biomimetic TME and provides new mechanistic insights into the effects of ADSC-released fatty acids on cancer cells as oncometabolites.