scholarly journals New Mechanical Fat Separation Technique: Adjustable Regenerative Adipose-tissue Transfer (ARAT) and Mechanical Stromal Cell Transfer (MEST)

2020 ◽  
Vol 2 (4) ◽  
Author(s):  
H Eray Copcu ◽  
Sule Oztan

Abstract Background Adipose tissue is not only a very important source of filler but also the body’s greatest source of regenerative cells. Objectives In this study, adipose tissue was cut to the desired dimensions using ultra-sharp blade systems to avoid excessive blunt pressure and applied to various anatomical areas—a procedure known as adjustable regenerative adipose-tissue transfer (ARAT). Mechanical stromal cell transfer (MEST) of regenerative cells from fat tissue was also examined. Methods ARAT, MEST, or a combination of these was applied in the facial area of a total of 24 patients who were followed for at least 24 months. The integrity of the fat tissue cut with different diameter blades is shown histopathologically. The number and viability of the stromal cells obtained were evaluated and secretome analyses were performed. Patient and surgeon satisfaction were assessed with a visual analog scale. Results With the ARAT technique, the desired size fat grafts were obtained between 4000- and 200-micron diameters and applied at varying depths to different aesthetic units of the face, and a guide was developed. In MEST, stromal cells were obtained from 100 mL of condensed fat using different indication-based protocols with 93% mean viability and cell counts of 28.66 to 88.88 × 106. Conclusions There are 2 main complications in fat grafting: visibility in thin skin and a low retention rate. The ARAT technique can be used to prevent these 2 complications. MEST, on the other hand, obtains a high rate of fat and viable stromal cells without applying excessive blunt pressure. Level of Evidence: 4

2022 ◽  
Vol 8 ◽  
pp. 205951312110478
Author(s):  
H Eray Copcu

Background Regenerative medicine is the fastest developing branch of plastic surgery in recent times. Adipose tissue is one of the largest and most important sources in the body for stromal cells. Although mechanical isolation methods are both very popular and have many advantages, they still have no accepted protocols. Objective We developed new protocols called indication-based protocols (IPs) for standardization and new techniques called mechanical stromal-cell transfer (MEST) by using ultra-sharp blades and dilution of adipose tissue with different solutions (saline, Ringer and 5% Dextrose) Methods & material: In order to obtain the desired physical structure (liquid, gel, solid) and the desired volume, four different types of IPs have been defined. Adipose tissue was prediluted with different solutions using 10 or 20 cc injectors in IPs 1 and 2, while condensed adipose tissue was used directly in IPs 3 and 4. Results In MEST, stromal cells were obtained from 100 mL of condensed fat using different IPs with 92% mean viability and cell counts of 26.80–91.90 × 106. Stromal cells can be obtained in the desired form and number of cells by using four different IPs. Conclusion Isolation of stromal cells by cutting fat with sharp blades will prevent the death of fat tissue and stromal cells and will allow high viability and cell count with our new technique. Predilution with different solutions: Diluting the condensed adipose tissue with the desired solutions (saline, Ringer or 5% Dextrose) before the adinizing process will provide even more stromal cells. Lay Summary Obtaining regenerative stromal cells from adipose tissue can be done by two methods: Enzymatic and mechanical. Mechanical methods have many advantages. Although mechanical stromal cell extraction from adipose tissue is very popular and many techniques have been described, there are still no accepted protocols, definition for the end product, and no consensus on the status of the stromal cells. In this study, stromal cells were obtained mechanically by using ultra-sharp blade systems, without exposing adipose tissue to blunt trauma. Thus, a higher number of cells and higher viability could be obtained. An “Indication based” protocol has been defined for the first time in order to obtain the desired number and status (solid, semi-solid, liquid) end product. Diluting the condensed adipose tissue with the desired solutions (saline, Ringer or 5% Dextrose) before the adinizing process will provide even more stromal cells. This will provide an opportunity for clinicians to obtain and apply a stromal cell solution for different indications in different anatomical regions.


Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1689-1689
Author(s):  
Jichun Chen ◽  
Neal S. Young

Abstract Immune-mediated bone marrow (BM) failure has been modeled in the mouse by infusion of lymph node cells from allogeneic C57BL/6 (B6) donors into major or minor histocompatibility antigen-mismatched recipients (Chen et al., Blood 2004; Bloom et al., Exp Hematol 2004, Chen et al., J Immunol 2007). Co-infusion of limited numbers of CD4+CD25+ regulatory T lymphocytes (Tregs) can alleviate clinical manifestations by suppressing the expansion of pathogenic T cells (Chen et al., J Immunol 2007). In the current study, we investigated the effectiveness of Tregs and suppressor cells contained in BM stroma in this fatal disease. Infusion of fewer than 3 × 103 Tregs to each recipient mouse had only a minor effect in preserving BM cells and did not prevent pancytopenia. Fifteen-50 × 103 thymic Tregs was moderately protective: blood WBC, RBC, platelet and BM cell counts at three weeks after cell infusion were 197%, 116%, 155% and 158% of those of control animals that did not receive Treg infusion; 5–10 × 103 B6 splenic Tregs produced the largest effect as WBC, RBC, platelet and BM cell counts were 275%, 143%, 276%, and 198% of controls. Overall, Treg therapy was helpful but its effectiveness was limited and variable among individual recipients as no antigen-specific Tregs can be identified for the treatment of BM failure. Learned about the immunosuppressive effects of mesenchymal stem cells (MSCs), we went on to test the effectiveness of stromal cells as another therapeutic modality for BM failure, since stromal cells contain MSCs. These cells were derived from B6 BM by culture in α-modified Eagle medium at 33°C with 5% CO2 for two weeks. After separating the non-adherent cells, we detached the adherent stromal cells and infused them into TBI + B6 LN-infused C.B10 mice. Injection of 106 stromal cells at the time of LN cell infusion effectively preserved WBCs (3.09 ± 0.51 vs 0.61 ± 0.18), RBCs (8.72 ± 0.14 vs 3.52 ± 0.46), platelets (924 ± 93 vs 147 ± 25) and BM cells (186.6 ± 8.7 vs 52.7 ± 7.8) when compared to LN-cell-infused mice without stromal cell addition. Delayed stromal cell injection at day 9 after LN cell infusion had only a mild effect on the preservation of RBCs (147%), platelets (276%) and BM cells (223%) and no effect on WBCs (64%), and infusion of non-adherent cells from the same stromal cell culture had no therapeutic effect. Stromal cell-infused mice had higher proportion of FoxP3+CD4+ cells in the peripheral blood (59.7 ± 10.7% vs 29.8 ± 5.4%) and more Lin−CD117+CD34− hematopoietic stem and progenitor cells in the BM (591 ± 95 vs 60 ± 43, thousand) in comparison to LN cell infused mice without stromal cell treatment. Mitigation of pathogenic T cells, including both CD4 and CD8 T lymphocytes, is the potential mechanism for the effectiveness of Treg and stromal cell therapies that helped to protect hematopoietic stem and progenitor cells in the BM of affected animals. Figure Figure


Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 863 ◽  
Author(s):  
Kristin Eckel-Mahan ◽  
Aleix Ribas Latre ◽  
Mikhail G. Kolonin

Adipose tissue (AT) is comprised of a diverse number of cell types, including adipocytes, stromal cells, endothelial cells, and infiltrating leukocytes. Adipose stromal cells (ASCs) are a mixed population containing adipose progenitor cells (APCs) as well as fibro-inflammatory precursors and cells supporting the vasculature. There is growing evidence that the ability of ASCs to renew and undergo adipogenesis into new, healthy adipocytes is a hallmark of healthy fat, preventing disease-inducing adipocyte hypertrophy and the spillover of lipids into other organs, such as the liver and muscles. However, there is building evidence indicating that the ability for ASCs to self-renew is not infinite. With rates of ASC proliferation and adipogenesis tightly controlled by diet and the circadian clock, the capacity to maintain healthy AT via the generation of new, healthy adipocytes appears to be tightly regulated. Here, we review the contributions of ASCs to the maintenance of distinct adipocyte pools as well as pathogenic fibroblasts in cancer and fibrosis. We also discuss aging and diet-induced obesity as factors that might lead to ASC senescence, and the consequences for metabolic health.


Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1764-1764
Author(s):  
Hendrik W van Deventer ◽  
Todd Hoffert ◽  
Michelle L West ◽  
Qing Ping Wu ◽  
Jonathan S Serody

Abstract Abstract 1764 Background: Chemotherapy resistance in chronic lymphocytic leukemia (CLL) is in part mediated by anti-apoptotic signals produced by bone marrow stromal cells. Identifying these signals is the first step to overcoming this resistance. ERDR1 was initially described as an inducer of hemoglobinization. We now present evidence that it inhibits apoptosis of CLL cells. Methods/Results: Previously, we showed that wild type (WT) but not CCR5−/− mesenchymal cells increase pulmonary melanoma metastasis in CCR5−/− mice. This observation led us to compare gene expression in the lungs of these mice. Using an Affymetrix expression array, we found Erdr1 was differentially expressed in the wild type mice. To show that the increase in metastasis was mediated by Erdr1, we transferred WT pulmonary mesenchymal cells transfected with Erdr1 shRNA or non-targeted control shRNA. CCR5−/− mice receiving Erdr1 knockdown mesenchymal cells had a 27.3% to 37% decrease in metastasis compared to animals receiving control transfected cells (p<0.01). One explanation for the decrease in metastasis would be a failure of the Erdr1 knockdown cells to survive in the lung. Since the knockdown and control vectors express EGFP, we were able to compare the quantity of transfected cells surviving in the lung by applying an EGFP ELISA to lung homogenates. These experiments showed no difference in the number of surviving mesenchymal cells. These results suggested Erdr1 was acting as a pro-survival factor for the melanoma cells. Since ERDR1 expression is the highest in the bone marrow, we compared the survival of CLL cells co-cultured with control and Erdr1 knockdown cells. In these experiments, stable Erdr1 knockdown and control clones were selected after the transfection of the bone marrow stromal cell line M2-10B4. Peripheral blood samples were then collected from 10 untreated CLL patients and co-cultured with these stromal cell lines. After 72 and 96 hours, total cell counts and apoptosis were measured using Annexin V/PI. At both time points, the cell counts were higher when the CLL cells were co-cultured with control cell lines compared to Erdr1 knockdown lines (OR 1.88 ± 0.27, 2.52 ± 0.66 respectively). The increase in total cell number was associated with a decrease in the percentage of apoptotic cells (OR 0.69 ± 0.18, 0.58 ± 0.12 respectively). Since Erdr1 was differentially expressed in WT compared to CCR5−/− mice, we considered the regulation of this gene by chemokine agonists. In these experiments, 100 ng/ml of CCL4 was added to WT and CCR5−/− PMCs and mRNA was harvested at 12, 24, and 48 hours. Using real-time PCR, we found that Erdr1 expression was increased compared to baseline in the WT mesenchymal cells by 1.33 fold ± 0.06 (p < 0.05) after 24 hours. By 48 hours, expression had increased by 3.36 fold ± 0.14 (p < 0.001). As expected, CCL4 did not increase expression of Erdr1 in CCR5−/− mesenchymal cells. Since Erdr1 is associated with hemoglobinization, we also investigated the effect of hypoxia on Erdr1 expression. In these experiments, deferoxamine was added to the stromal cell line M2-10B4 at varying concentrations. Significant fold increases in Erdr1 expression were seen after 48 hours of 1.43 ± 0.10 (50nM), 1.96 ± 0.08, (100nM), and 2.44 ± 0.01 (200nM). Implications for the treatment of human disease: Other investigators have shown that stromal cells such as nurse-like cells and CLL cells can interact to induce CCL4 and promote CLL cell survival. Our work suggests a novel pathway by which this may take place. Disclosures: No relevant conflicts of interest to declare.


2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Wang Jin ◽  
Yiming Zhao ◽  
Yonghao Hu ◽  
Dingding Yu ◽  
Xiang Li ◽  
...  

Endometrial regenerative cells (ERCs) are mesenchymal-like stromal cells obtained from human menstrual blood, whose positive therapeutic effects have been validated in several experimental models. Stromal cell-derived factor-1 (SDF-1), the ligand for CXCR4, plays an important role in the migration of mesenchymal stromal cells. The purpose of this study was to investigate the role of the SDF-1/CXCR4 pathway in the therapeutic effects of ERCs in a mouse sepsis model. Through preexperiment and confirmation, wild-type C57BL/6 mice were intraperitoneally injected with 10 mg/kg lipopolysaccharide (LPS). The therapeutic effects of ERCs with different pretreatments were evaluated by assessing sepsis-related symptoms, detecting tissue damage and measuring levels of inflammatory and oxidative stress-related factors. The in vitro experiments demonstrated that there was a much higher CXCR4 expression on ERCs when they were cocultured with SDF-1. The ex vivo experiment results showed that SDF-1 expression significantly increased in mouse tissues. Further experiments also confirmed that, compared with the unmodified ERC treatment group, SDF-1 pretreatment significantly enhanced the therapeutic effects of ERCs on alleviating sepsis symptoms, ameliorating pathological changes, reducing Bax level, and increasing Bcl-2 and PCNA expressions in mouse liver tissues. Furthermore, it was also found that SDF-1-pretreated ERCs contributed to reducing the levels of proinflammatory cytokines (TNF-α, IL-1β) and increasing the levels of anti-inflammatory factors (IL-4, IL10) in mouse serum, liver, and lung. Moreover, SDF-1-pretreated ERCs could also significantly decrease the levels of iNOS and MDA and increase the expression of Nrf2, HO-1, and SOD in liver tissues. Taken together, these results indicate that SDF-1 pretreatment plays a key role in improving the therapeutic effects of ERCs in alleviating sepsis-related symptoms, reducing tissue damage, regulating inflammatory imbalance, and relieving oxidative stress in a mouse sepsis model, which provides more possibilities for the clinical application of ERCs in sepsis and relevant diseases.


2019 ◽  
Vol 216 (9) ◽  
pp. 1999-2009 ◽  
Author(s):  
Batika M.J. Rana ◽  
Eric Jou ◽  
Jillian L. Barlow ◽  
Noe Rodriguez-Rodriguez ◽  
Jennifer A. Walker ◽  
...  

Group-2 innate lymphoid cells (ILC2), type-2 cytokines, and eosinophils have all been implicated in sustaining adipose tissue homeostasis. However, the interplay between the stroma and adipose-resident immune cells is less well understood. We identify that white adipose tissue–resident multipotent stromal cells (WAT-MSCs) can act as a reservoir for IL-33, especially after cell stress, but also provide additional signals for sustaining ILC2. Indeed, we demonstrate that WAT-MSCs also support ICAM-1–mediated proliferation and activation of LFA-1–expressing ILC2s. Consequently, ILC2-derived IL-4 and IL-13 feed back to induce eotaxin secretion from WAT-MSCs, supporting eosinophil recruitment. Thus, MSCs provide a niche for multifaceted dialogue with ILC2 to sustain a type-2 immune environment in WAT.


2014 ◽  
Vol 26 (03) ◽  
pp. 1450045
Author(s):  
Shou-Cheng Teng ◽  
Li-Ting Li ◽  
Shyi-Gen Chen ◽  
Tien-Mu Chen ◽  
Cheng-Hao Liao ◽  
...  

Adipose tissue is not only an ideal material for soft tissue filling and augmentation, but also a plentiful source of regenerative cells in adipose-derived stromal vascular fraction. Enriched fat tissue with adipose-derived stromal vascular fraction cells (ADSVFCs) can improve the fat graft survival, as shown in previous studies. The objective of this study is to estimate the effects of various dosages of ADSVFCs on fat grafts. We hypothesized that the improvement of ADSVFCs on fat transplantation quality would be dose dependent. Fat tissues were obtained by liposuction surgery. ADSVFCs were mixed into fat tissue, and the ADSVFC-enriched fat tissues were implanted subcutaneously into nude mice. The histological findings showed that the neoangiogenesis and integrity of grafted fat cells significantly increased with increasing dosages of ADSVFCs. Higher dosages of ADSVFCs also significantly reduced the side effects of the fat grafts, including reducing inflammation, cell infiltration, fibrosis and cyst formation. In conclusion, ADSVFC supplemented fat implants can improve angiogenesis and anti-inflammation. However, these effects of ADSVFCs on fat graft quality are dose dependent.


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