Application Of Adipose-derived Mesenchymal Stem Cells Research Articles

Application of adipose-derived stem cells in ischemic heart disease: theory, potency, and advantage.

Adipose-derived mesenchymal stem cells (ASCs) represent an innovative candidate to treat ischemic heart disease (IHD) due to their abundance, renewable sources, minor invasiveness to obtain, and no ethical limitations. Compared with other mesenchymal stem cells, ASCs have demonstrated great advantages, especially in the commercialization of stem cell-based therapy. Mechanistically, ASCs exert a cardioprotective effect not only through differentiation into functional cells but also via robust paracrine of various bioactive factors that promote angiogenesis and immunomodulation. Exosomes from ASCs also play an indispensable role in this process. However, due to the distinct biological functions of ASCs from different origins or donors with varing health statuses (such as aging, diabetes, or atherosclerosis), the heterogeneity of ASCs deserves more attention. This prompts scientists to select optimal donors for clinical applications. In addition, to overcome the primary obstacle of poor retention and low survival after transplantation, a variety of studies have been dedicated to the engineering of ASCs with biomaterials. Besides, clinical trials have confirmed the safety and efficacy of ASCs therapy in the context of heart failure or myocardial infarction. This article reviews the theory, efficacy, and advantages of ASCs-based therapy, the factors affecting ASCs function, heterogeneity, engineering strategies and clinical application of ASCs.

Current applications of adipose-derived mesenchymal stem cells in bone repair and regeneration: A review of cell experiments, animal models, and clinical trials.

In the field of orthopaedics, bone defects caused by severe trauma, infection, tumor resection, and skeletal abnormalities are very common. However, due to the lengthy and painful process of related surgery, people intend to shorten the recovery period and reduce the risk of rejection; as a result, more attention is being paid to bone regeneration with mesenchymal stromal cells, one of which is the adipose-derived mesenchymal stem cells (ASCs) from adipose tissue. After continuous subculture and cryopreservation, ASCs still have the potential for multidirectional differentiation. They can be implanted in the human body to promote bone repair after induction in vitro, solve the problems of scarce sources and large damage, and are expected to be used in the treatment of bone defects and non-union fractures. However, the diversity of its differentiation lineage and the lack of bone formation potential limit its current applications in bone disease. Here, we concluded the current applications of ASCs in bone repair, especially with the combination and use of physical and biological methods. ASCs alone have been proved to contribute to the repair of bone damage in vivo and in vitro. Attaching to bone scaffolds or adding bioactive molecules can enhance the formation of the bone matrix. Moreover, we further evaluated the efficiency of ASC-committed differentiation in the bone in conditions of cell experiments, animal models, and clinical trials. The results show that ASCs in combination with synthetic bone grafts and biomaterials may affect the regeneration, augmentation, and vascularization of bone defects on bone healing. The specific conclusion of different materials applied with ASCs may vary. It has been confirmed to benefit osteogenesis by regulating osteogenic signaling pathways and gene transduction. Exosomes secreted by ASCs also play an important role in osteogenesis. This review will illustrate the understanding of scientists and clinicians of the enormous promise of ASCs’ current applications and future development in bone repair and regeneration, and provide an incentive for superior employment of such strategies.

Comprehensive RNA expression profile of therapeutic adipose‑derived mesenchymal stem cells co‑cultured with degenerative nucleus pulposus cells.

Stem cell-based therapy is a promising alternative to conventional approaches to treating intervertebral disc degeneration (IDD). However, comprehensive understanding of stem cell-based therapy at the gene level is still lacking. In the present study, we identified the expression profiles of messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) expressed within a co-culture system of adipose-derived mesenchymal stem cells (ASCs) and degenerative nucleus pulposus cells (NPCs) and explored the signaling pathways involved and their regulatory networks. Microarray analysis was used to compare ASCs co-cultured with degenerative NPCs to ASCs cultured alone, and the underlying regulatory pattern, including the signaling pathways and competing endogenous RNA (ceRNA) network, was analyzed with robust bioinformatics methods. The results showed that 360 lncRNAs and 1757 mRNAs were differentially expressed by ASCs, and the microarray results were confirmed by quantitative PCR. Moreover, 589 Gene Ontology terms were upregulated, whereas 661 terms were downregulated. A total of 299 signaling pathways were significantly altered. A Path-net and a Signal-net were built to show interactions among differentially expressed genes. An mRNA-lncRNA co-expression network was constructed to reveal the interplay among differentially expressed mRNAs and lncRNAs, whereas a ceRNA network was built to investigate their connections with microRNAs involved in IDD. To the best of our knowledge, this original and comprehensive exploration reveals differentially expressed lncRNAs and mRNAs of ASCs stimulated by degenerative NPCs, underscoring the regulation pattern within the co-culture system at the gene level. These data may further understanding of NPC-directed differentiation of ASCs and facilitate the application of ASCs in future treatments for IDD.

Application of mesenchymal stem cells to enhance non-union bone fracture healing.

ECM components include a number of osteoinductive and osteoconductive factors, which are involved in bone fracture healing. In this study, a combination of adipose derived mesenchymal stem cells (Ad-MSCs), cancellous bone graft (CBG), and chitosan hydrogel (CHI) was applied to the non-union bone fracture and healing effects were evaluated for the first time. After creation of animal models with non-union fracture in rats, they were randomly classified into seven groups. Radiography at 0, 2, 4, and 8 weeks after surgery, indicated the positive effects of Ad-MSCs + CBG + CHI and Ad-MSCs + CBG in treatment of bone fractures as early as 2 weeks after the surgery. These data were confirmed with both biomechanical and histological studies. Gene expression analyses of Vegf and Bmp2 showed a positive effect of Ad-MSCs on vascularization and osteogenic differentiation in all groups receiving Ad-MSCs, as shown by real-time PCR. Immunofluorescence analysis and RT-PCR results indicated existence of human Ad-MSCs in the fractured region 8 weeks post-surgery. In conclusion, we suggest that application of Ad-MSCs, CBG, and CHI, could be a suitable combination for osteoinduction and osteoconduction to improve non-union bone fracture healing. Further investigations are required to determine the exact mechanisms involved in this process before moving to clinical studies. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 301-311, 2019.

Injured Achilles Tendons Treated with Adipose-Derived Stem Cells Transplantation and GDF-5.

Tendon injuries represent a clinical challenge in regenerative medicine because their natural repair process is complex and inefficient. The high incidence of tendon injuries is frequently associated with sports practice, aging, tendinopathies, hypertension, diabetes mellitus, and the use of corticosteroids. The growing interest of scientists in using adipose-derived mesenchymal stem cells (ADMSC) in repair processes seems to be mostly due to their paracrine and immunomodulatory effects in stimulating specific cellular events. ADMSC activity can be influenced by GDF-5, which has been successfully used to drive tenogenic differentiation of ADMSC in vitro. Thus, we hypothesized that the application of ADMSC in isolation or in association with GDF-5 could improve Achilles tendon repair through the regulation of important remodeling genes expression. Lewis rats had tendons distributed in four groups: Transected (T), transected and treated with ADMSC (ASC) or GDF-5 (GDF5), or with both (ASC+GDF5). In the characterization of cells before application, ADMSC expressed the positive surface markers, CD90 (90%) and CD105 (95%), and the negative marker, CD45 (7%). ADMSC were also differentiated in chondrocytes, osteoblast, and adipocytes. On the 14th day after the tendon injury, GFP-ADMSC were observed in the transected region of tendons in the ASC and ASC+GDF5 groups, and exhibited and/or stimulated a similar genes expression profile when compared to the in vitro assay. ADMSC up-regulated Lox, Dcn, and Tgfb1 genes expression in comparison to T and ASC+GDF5 groups, which contributed to a lower proteoglycans arrangement, and to a higher collagen fiber organization and tendon biomechanics in the ASC group. The application of ADMSC in association with GDF-5 down-regulated Dcn, Gdf5, Lox, Tgfb1, Mmp2, and Timp2 genes expression, which contributed to a lower hydroxyproline concentration, lower collagen fiber organization, and to an improvement of the rats’ gait 24 h after the injury. In conclusion, although the literature describes the benefic effect of GDF-5 for the tendon healing process, our results show that its application, isolated or associated with ADMSC, cannot improve the repair process of partial transected tendons, indicating the higher effectiveness of the application of ADMSC in injured Achilles tendons. Our results show that the application of ADMSC in injured Achilles tendons was more effective in relation to its association with GDF-5.

Local application of adipose-derived mesenchymal stem cells supports the healing of fistula: prospective randomised study on rat model of fistulising Crohn’s disease

Objective: Local application of adipose-derived mesenchymal stem cells (ADSC) represents a novel approach for the management of perianal fistula in patients with Crohn’s disease. A randomised study on an animal model was performed to investigate the efficacy and to detect the distribution of implanted ADSCs by bioluminescence (BLI).Materials and methods: A caecostomy was used as a fistula model in 32 Lewis rats. The ADSCs were isolated from transgenic donor expressing firefly luciferase. Animals were randomly assigned to groups given injections of 4 × 106 cells (n = 16, group A) or placebo (n = 16, group B) in the perifistular tissue. Fistula drainage assessment was used to evaluate the fistula healing. After application of D-luciferin, cell viability and distribution was detected using an IVIS Lumina XR camera on days 0, 2, 7, 14 and 30.Results: The fistula was identified as healed in 6 (38%) animals in group A vs. 1 case (6.3%) in group B (p = .033). The BLI was strongest immediately after administration of ADSCs 31.2 × 104 (6.09–111 × 104) p/s/cm2/sr. The fastest decrease was observed within the first 2 days when values fell by 50.2%. The BLI 30 days after injection was significantly higher in animals with healed fistulas – 8.23 × 104 (1.18–16.9 × 104) vs. 1.74 × 104 (0.156–6.88 × 104); p = .0393.Conclusions: Local application of ADSCs resulted in significantly higher fistula closure rate on an animal model. BLI monitoring was proved to be feasible and showed rapid reduction of the ADSC mass after application. More viable cells were detected in animals with healed fistula at the end of the follow-up.

Evaluation of Serum-Free, Xeno-Free Cryopreservation Solutions for Human Adipose-Derived Mesenchymal Stem Cells.

Adipose-derived mesenchymal stem cells (ASCs) have the potential to differentiate into cells of mesodermal origin, such as osteoblasts, adipocytes, myocytes, and chondrocytes, and cryopreservation is currently performed as a routine method for preserving ASCs to safely acquire large numbers of cells. For clinical application of ASCs, serum-free, xeno-free cryopreservation solutions should be used. This study determined the viability and adipo-osteogenic potential of cryopreserved ASCs using four cryopreservation solutions: 10% DMSO, Cell Banker 2 (serum free), Stem Cell Banker (=Cell Banker 3: serum free, xeno free), and TC protector (serum free, xeno free). The viability of the cryopreserved ASCs was over 80% with all cryopreservation solutions. No difference in the adipo-osteogenic potential was found between the cells that did or did not undergo cryopreservation in these cryopreservation solutions. These data suggest that Cell Banker 3 and TC protector are comparable with 10% DMSO and Cell Banker 2 for ASCs, and cryopreserved as well as noncryopreserved ASCs could be applied for regenerative medicine.

Mo1707 Serial Treatment With Anti-TNF Ab Reduces Tumor Burden in a Murine Model of Chronic Colitis, Compared to Single Dose Anti-TNF

Background: Local application of adipose-derived mesenchymal stem cells (ADSCs) is a promising method for the treatment of perianal fistula in patients with Crohn's disease. There is a lack of data evaluating the distribution and viability of ADSCs after application. Therefore more studies are needed including the technique of in vivo monitoring. Aim: The aim of the study was to evaluate the effect of ADSCs application on fistula healing regarding their viability and distribution in perifistular region. Methods: Coecostomy was used as a fistula model in Lewis rats. The inguinal adipose tissue was harvested from transgenic donor expressing firefly luciferase (LEW-Tg(Rosa-luc)11Jmsk; Jichi Medical School, Japan). The ADSCs were isolated using collagenase technique and injected (1-2*106 cells/ml) in perifistular tissue of 16 rats. After application of D-luciferin (25mg/kg), animals were imaged in IVIS Lumina XR camera on days 0, 2, 7, 14 and 30. Fistula drainage assessment (FDA) was used to evaluate the fistula healing. Animals from interventional group were compared to 30 rats with untreated fistula. Results: There was no morbidity or mortality after intervention. The fistula was identified as healed according to FDA in 6 (37.5%) animals after ADSCs injection vs. 1 case (3.3%) with spontaneous closure in control group (p< 0.0047). Bioluminescence was strongest 2 days after application (31.2*104 (6.10-11.0*104). The amount of viable cells remained stable during first 7 days, which confirms successful cells seating. The signal started to decrease slowly afterwards day 14: 6.4*104 (0.5-28.8*104) and day 30: 2.8*104 (0.16-16.9*104) (values are medians in p/s/cm2/sr) (fig.1). The luminescence 30 days after injection was significantly higher in animals with healed fistula 8.23*104 (1.18*1041.70*105) vs. 2.75*104 (4.93*103-6.88*104); p=0.18. Conclusion: Perifistular application of ADSCs and in vivo monitoring using bioluminescence are technically feasible and doesn't cause mortality or morbidity. Local treatment with ADSCs led to significantly more frequent fistula closure compared to the control group. The fistula healing was associated with significantly higher concentration of viable ADSCs in perifistular region, which was confirmed by stronger bioluminescence signal. Supported by NT13708

Mo1706 Local Application of Adipose-Derived Mesenchymal Stem Cells Can Lead to Intestinal Fistula Healing - Experimental Study

Background: Local application of adipose-derived mesenchymal stem cells (ADSCs) is a promising method for the treatment of perianal fistula in patients with Crohn's disease. There is a lack of data evaluating the distribution and viability of ADSCs after application. Therefore more studies are needed including the technique of in vivo monitoring. Aim: The aim of the study was to evaluate the effect of ADSCs application on fistula healing regarding their viability and distribution in perifistular region. Methods: Coecostomy was used as a fistula model in Lewis rats. The inguinal adipose tissue was harvested from transgenic donor expressing firefly luciferase (LEW-Tg(Rosa-luc)11Jmsk; Jichi Medical School, Japan). The ADSCs were isolated using collagenase technique and injected (1-2*106 cells/ml) in perifistular tissue of 16 rats. After application of D-luciferin (25mg/kg), animals were imaged in IVIS Lumina XR camera on days 0, 2, 7, 14 and 30. Fistula drainage assessment (FDA) was used to evaluate the fistula healing. Animals from interventional group were compared to 30 rats with untreated fistula. Results: There was no morbidity or mortality after intervention. The fistula was identified as healed according to FDA in 6 (37.5%) animals after ADSCs injection vs. 1 case (3.3%) with spontaneous closure in control group (p< 0.0047). Bioluminescence was strongest 2 days after application (31.2*104 (6.10-11.0*104). The amount of viable cells remained stable during first 7 days, which confirms successful cells seating. The signal started to decrease slowly afterwards day 14: 6.4*104 (0.5-28.8*104) and day 30: 2.8*104 (0.16-16.9*104) (values are medians in p/s/cm2/sr) (fig.1). The luminescence 30 days after injection was significantly higher in animals with healed fistula 8.23*104 (1.18*1041.70*105) vs. 2.75*104 (4.93*103-6.88*104); p=0.18. Conclusion: Perifistular application of ADSCs and in vivo monitoring using bioluminescence are technically feasible and doesn't cause mortality or morbidity. Local treatment with ADSCs led to significantly more frequent fistula closure compared to the control group. The fistula healing was associated with significantly higher concentration of viable ADSCs in perifistular region, which was confirmed by stronger bioluminescence signal. Supported by NT13708