Source Of Adipose-derived Stem Cells Research Articles

Comparison of the infrapatellar and subcutaneous adipose tissue biopsy material as a source of mesenchymal stromal cells for regenerative medicine in traumatology

Objective. The use of infrapatellar fat pad (IPFP) as the source of mesenchymal stem cells (MSCs), show an age-independent proliferation and differentiation potential. In addition, the pronounced chondrogenic potential of IPFP-ASCs makes them promising candidates for research for use in other methodsof regenerative therapy. Methods. A direct immunohistochemical study was carried out in serial paraffin sections of the biopsy material of Hoffa`s fat pad and subcutaneus fat tissue, using monoclonal antibodies. The minimum criteria established by the International Society for Cell Therapy to ensure the identity of MSCs use CD73 and CD105 as positive markers and CD34, CD31, CD45 as a negative. Results. According to the results of histological, immunohistochemical, morphometric and statistical studies, it was found that in the biopsy of both tissues the relative number of cells with an immunoprofile CD105+, CD73+, CD34–, CD31–, CD45– in the standard field of view (×200) was: 7.25 (5.42; 8.89 ) %, and 11.11 (8.46; 13.45) %, no statistically significant difference was found between comparison groups (р > 0,05). Conclusion. The therapeutic effect of mesenchymal stem cells of subcutaneous adipose tissue has been proven, thus the identification of similar cells in the biopsy material and relatively similar density in the Hoffa body makesit an important source of adipose-derived stem cells that can be used for regenerative engineering tissue.

Cadaveric Adipose-Derived Stem Cells for Regenerative Medicine and Research.

Advances in regenerative medicine have enabled the search for new solutions to current health problems in so far unexplored fields. Thus, we focused on cadaveric subcutaneous fat as a promising source of adipose-derived stem cells (ADSCs) that have potential to differentiate into different cell lines. With this aim, we isolated and characterized ADSCs from cadaveric samples with a postmortem interval ranging from 30 to 55 h and evaluated their ability to differentiate into chondrocytes or osteocytes. A commercial ADSC line was used as reference. Morphological and protein expression analyses were used to confirm the final stage of differentiation. Eight out of fourteen samples from patients were suitable to complete the whole protocol. Cadaveric ADSCs exhibited features of stem cells based upon several markers: CD29 (84.49 ± 14.07%), CD105 (94.38 ± 2.09%), and CD44 (99.77 ± 0.32%). The multiparametric assessment of differentiation confirmed the generation of stable lines of chondrocytes and osteocytes. In conclusion, we provide evidence supporting the feasibility of obtaining viable postmortem human subcutaneous fat ADSCs with potential application in tissue engineering and research fields.

Ultrasound cavitation: a reliable non-enzymatic method for adipose-derived mesenchymal stem cell (ADSC) isolation

BackgroundAdipose tissue is known to serve as an abundant and readily accessible source of adipose-derived stem cells (ADSCs) as an alternative to bone marrow. Collagenase is one of the most widely used methods for the isolation of ADSCs from adipose tissue, but it takes a long time, and there are also debates about safety. We propose an ultrasonic cavitation-treated method that can significantly reduce time and avoid the problem of using xenogeneic enzymes in ADSCs isolation.MethodsADSCs were isolated from adipose tissue using the enzyme treatment method and the ultrasonic cavitation treatment method. Cell proliferation was measured using cell viability assay. The expression levels of the surface markers of ADSCs were estimated by real-time PCR. After, ADSCs were cultured in chondrogenic, osteogenic, or adipogenic differentiation medium; the differentiation potential of ADCSs was analyzed by Alcian blue, Alizarin Red S, Oil Red O, and real-time PCR.ResultsThe cells treated with collagenase and ultrasound had similar cell yields and proliferation after isolation. The difference in the expression of surface markers of ADSCs was not statistically significant. ADSCs showed differentiation potential into adipocytes, osteocytes, and chondrocytes, and there was no difference between the enzyme treatment method and the ultrasonic cavitation treatment method. The yield of the ADSC increased in time- and intensity dependently.ConclusionsUltrasound certainly serves as a promising method in advancing ADSC isolation technology.

Use of adipose derived stem cells in Treacher Collins syndrome.

Treacher Collins syndrome (TCS) is a rare congenital disorder of craniofacial development. TCS occurs with an incidence of 1:50,000, and more than 60% of TCS cases have no previous family history and arise as the result of de novo mutations. The high rate of de novo mutations, together with the extreme variability in the degree to which individuals can be affected, makes the provision of genetic counseling extremely complicated. Consequently, every case of TCS is unique and needs to be assessed individually. Patients with TCS frequently undergo multiple reconstructive surgeries from birth through adulthood, which rarely are fully corrective in the long-term. The nascent field of regenerative medicine offers the promise to improve some of these treatments. In particular, structural fat grafting (SFG) seems to be a good strategy not only to restore the normal volume and contour of the face, but also to provide a source of adipose-derived stem cells (ADSCs) with a multilineage differentiation potential. In this work, we present genetical analyses of ADSC affected by TCS. ADSCs from were analyzed for their stemness properties and shared many characteristics with those of a healthy subject. Screening of the genome of the TCS patient using array-Comparative Genomic Hybridization allowed us to identify some chromosomal imbalances that are probably associated with TCS. We found that some alterations, involving the TIMELESS gene, were usually associated with embryonic stem cells. With the aim to improve the final results, we need to consider combining knowledge of genetic alterations and expression profiles as a fundamental step before starting with surgical procedures.

Purification and characterization of human adipose-resident microvascular endothelial progenitor cells

Human adipose tissue is a rich source of adipose-derived stem cells (ASCs) and vascular endothelial progenitor cells (EPCs). However, no standardized method has been established for the isolation and purification of adipose-derived EPCs (AEPCs). The aim of this study was to establish a method for the isolation and purification of AEPCs. The stromal vascular fraction (SVF) was extracted from human lipoaspirates, and the CD45−CD31+ fraction of the SVF was collected by magnetic-activated cell sorting (MACS). The CD45−CD31+ fraction was cultured for 4.5 days, followed by a second MACS separation to collect the CD31+ fraction. Purified AEPCs were expanded without being overwhelmed by proliferating ASCs, indicating that a high level (> 95%) of AEPC purification is a key factor for their successful isolation and expansion. AEPCs exhibited typical endothelial markers, including CD31, von Willebrand factor, and the isolectin-B4 binding capacity. AEPCs formed colonies, comparable to cultured human umbilical vein endothelial cells (HUVECs). Both AEPCs and HUVECs formed capillary-like networks in the tube formation assay, with no significant difference in network lengths. We are the first to establish a purification and expansion method to isolate these cells. Because adipose tissue is a clinically accessible and abundant tissue, AEPCs may have potential advantages as a therapeutic tool for regenerative medicine.

Adipose Tissue: Understanding the Heterogeneity of Stem Cells for Regenerative Medicine.

Adipose-derived stem cells (ASCs) have been increasingly used as a versatile source of mesenchymal stem cells (MSCs) for diverse clinical investigations. However, their applications often become complicated due to heterogeneity arising from various factors. Cellular heterogeneity can occur due to: (i) nomenclature and criteria for definition; (ii) adipose tissue depots (e.g., subcutaneous fat, visceral fat) from which ASCs are isolated; (iii) donor and inter-subject variation (age, body mass index, gender, and disease state); (iv) species difference; and (v) study design (in vivo versus in vitro) and tools used (e.g., antibody isolation and culture conditions). There are also actual differences in resident cell types that exhibit ASC/MSC characteristics. Multilineage-differentiating stress-enduring (Muse) cells and dedifferentiated fat (DFAT) cells have been reported as an alternative or derivative source of ASCs for application in regenerative medicine. In this review, we discuss these factors that contribute to the heterogeneity of human ASCs in detail, and what should be taken into consideration for overcoming challenges associated with such heterogeneity in the clinical use of ASCs. Attempts to understand, define, and standardize cellular heterogeneity are important in supporting therapeutic strategies and regulatory considerations for the use of ASCs.

New Trends In Treacher Collins Syndrome: Bony Reconstruction And Regenerative Therapy

Aim:Treacher Collins syndrome is a rare congenital disorder of craniofacial development with a highly variable pheonotype. This syndrome occurs with an incidence of 1:50,000, and more than 60% of the cases have no previous family history and arise as the result of de novo mutations. The disorder displays an intricate underlying dysmorphology. Affected patients may suffer life-threatening airway complications and functional difficulties involving sight, hearing, speech, and feeding. Deformation of facial structures produces a characteristic appearance that includes malar-zygomatic hypoplasia, periorbital soft tissue anomalies, maxillomandibular hypoplasia, and ear anomalies. Management requires a specialized craniofacial team, as comprehensive care starts at birth and may require life-long follow-up. Standard craniofacial procedures for bony and soft tissue reconstruction are used. This article outlines current treatment strategies and future concepts for surgical and regenerative management. Methods:The new field of regenerative medicine and therapy offers the promise to improve some of these treatments. In particular, Structural Fat Grafting (lipostructure) seems to be a good strategy to restore the normal volume and contour of the face, and to provide a source of adipose-derived stem cells (ADSCs) with a multilineage differentiation potential. In this work, we present the case of a young girl with Treacher Collins Syndrome who underwent serial sessions of fat grafting in addition to other surgical bony reconstructive techniques. ADSCs have been isolated from the patient’s lipoaspirate, and compared for their stemness properties with those of a healthy subject. Conclusion:Screening of the genome of the Treacher Collins patient using array-Comparative Genomic Hybridization (array-CGH) allowed us to identify some chromosomal imbalances that are probably associated with the syndrome.Correction of these imbalances and asymmetries by modulating ADSCs could be an innovative approach to improve and stabilize the results of the surgical treatment of Treacher Collin Syndrome.

Treatment of “En Coupe De Sabre “ Linear Scleroderma. Soft Tissue Augmentation and Regeneration by Autologous Fat Transfer

Aim: Frontal linear scleroderma, also known as ‘‘en coup de sabre,’’ is a congenital deformity characterized by atrophy and furrowing of the skin of the front parietal area above the level of the eyebrows. In most cases it occurs as a single paramedian line that may be associated with hypoplasia of underlying structures and facial hemiatrophy. In case of a wide lesion many reconstructive strategies have been proposed. The modern approach is aimed at augmentation of the tissue deficiency by using lipostructure and tissue regeneration. Autologous fat transfer (AFT) seems to be an effective strategy to restore the normal volume and contour of the face while providing a source of adipose-derived stem cells (ADSCs) with a multilineage differentiation potential. Methods: In this report, we present a rare case of linear scleroderma en coupe de sabre which was successfully managed with three stages of autologous fat grafting. Conclusion: The proposed uses for ADSCs in tissue repair and regeneration are quite impressive. Recent works on ADSCs would suggest that adult cells may prove to be an equally powerful regenerative tool in treating congenital and acquired maxillofacial disorders. More importantly, physicians, researchers and international associations need to cooperate in informing clinicians about what practices are based on evidence and to encourage support of additional research. There is increasing interest in a possible therapeutic effect of ADSCs from processed lipoaspirate for a wide spectrum of clinical applications in the facial and craniofacial area. AFT can be used in any facial area where soft tissue is lacking or where there is scarring, producing natural and long-lasting results. Mesenchymal stem cells represent a great tool in regenerative medicine. However, more definitive studies are needed to answer specific questions regarding the best technique to be used and the role of ADSCs. Autologous fat grafting provides a safe and easy approach for the treatment of linear scleroderma en coup de sabre, long-term clinically satisfactory results can be obtained.

Bisecting GlcNAc Protein N-Glycosylation Is Characteristic of Human Adipogenesis.

Human adipose tissue contains a major source of adipose-derived stem cells (ADSCs) that have the ability to differentiate into various cell types: in vitro, ADSCs can differentiate into mesenchymal lineages including adipocytes, while in vivo, ADSCs become mature adipocytes. Protein glycosylation has been shown to change in stem cell differentiation, and while ADSCs have been acknowledged for their therapeutic potential, little is known about protein glycosylation during human ADSC adipogenic differentiation. In the present study, the global membrane protein glycosylation of native adipocytes was compared to ADSCs from the same individuals as a model of in vivo adipogenesis. For in vitro adipogenesis, ADSCs were adipogenically differentiated in cell culture using an optimized, large-scale differentiation procedure. The membrane glycome of the differentiated ADSCs (dADSCs) was compared with mature adipocytes and the progenitor ADSCs. A total of 137 glycan structures were characterized across the three cell types using PGC-LC coupled with negative-ion electrospray ionization mass spectrometry (ESI-MS)/MS. Significantly higher levels of bisecting GlcNAc-type N-glycans were detected in mature adipocytes (32.1% of total glycans) and in in vitro dADSC progeny (1.9% of total glycans) compared to ADSCs. This was further correlated by the mRNA expression of the MGAT3 gene responsible for the enzymatic synthesis of this structural type. The bisecting GlcNAc structures were found on the majority of human native adipocyte membrane proteins, suggesting an important role in human adipocyte biology. Core fucosylation was also significantly increased during in vivo adipogenesis but did not correlate with an increase in Fut8 gene transcript. Unexpectedly, low abundance structures carrying rare β-linked Gal-Gal termini were also detected. Overall, the N-glycan profiles of the in vitro differentiated progeny did not reflect native adipocytes, and the results show that bisecting GlcNAc structures are a characteristic feature of human adipocyte membrane protein N-glycosylation. Raw MS files are available on GlycoPOST (ID: GPST000153 https://glycopost.glycosmos.org/).

Does Water-Jet Force Affect Cryopreserved Adipose-Derived Stem Cells? Evidence of Improved Cell Viability and Fat Graft Survival.

Adipose tissue harvested by liposuctions is an available source of adipose-derived stem cells (ASCs). Water-jet-assisted liposuction is a favorable method for fat collection with little mechanical damage. This study aimed to investigate whether or not the water-jet-assisted liposuction made a difference in the biological characteristics of cryopreserved ASCs and fat graft survival in cell-assisted lipotransfer. Human lipoaspirates were obtained from the abdomen or thighs of 20 female participants for body contouring. A single surgeon randomly harvested 50 mL of adipose tissue by the water-jet-assisted liposuction and the conventional liposuction, respectively. Adipose-derived stem cells were isolated from lipoaspirates and then cryopreserved for 4 weeks. Cryopreserved ASCs were used to examine the surface markers, cell proliferation, migration, and adipogenic differentiation in vitro. The fat survival of ASCs-enriched grafts from different liposuctions was measured in animal models. The cryopreserved ASCs with the water-jet assistance had better capacities of cell proliferation, migration, and adipogenic differentiation and achieved a better survival result of ASCs-enriched fat grafting. Cryopreservation of ASCs with the water-jet force showed more excellent biological characteristics. The water-jet-assisted liposuction was superior to the conventional liposuction in obtaining ASCs and fat survival of coimplantation with grafts.

Tuning Adipogenic Differentiation in ADSCs by Metformin and Vitamin D: Involvement of miRNAs.

Fat tissue represents an important source of adipose-derived stem cells (ADSCs), which can differentiate towards several phenotypes under certain stimuli. Definite molecules as vitamin D are able to influence stem cell fate, acting on the expression of specific genes. In addition, miRNAs are important modulating factors in obesity and numerous diseases. We previously identified specific conditioned media able to commit stem cells towards defined cellular phenotypes. In the present paper, we aimed at evaluating the role of metformin on ADSCs differentiation. In particular, ADSCs were cultured in a specific adipogenic conditioned medium (MD), in the presence of metformin, alone or in combination with vitamin D. Our results showed that the combination of the two compounds is able to counteract the appearance of an adipogenic phenotype, indicating a feedforward regulation on vitamin D metabolism by metformin, acting on CYP27B1 and CYP3A4. We then evaluated the role of specific epigenetic modulating genes and miRNAs in controlling stem cell adipogenesis. The combination of the two molecules was able to influence stem cell fate, by modulating the adipogenic phenotype, suggesting their possible application in clinical practice in counteracting uncontrolled lipogenesis and obesity-related diseases.

Characterization of Mitochondrial Bioenergetics of Adipose‐Derived Stem Cells from Burned and Non‐burned Patients

Prolonged hypermetabolism is a hallmark of severe burn injury (over 20% of the total body surface area) that produces cachexia and hinders the wound healing process. Topical treatment with adipose‐derived stem cells (ASCs) have been proposed as an attractive solution to hasten the recovery of burn wounds. A potentially viable source of ASC is from the subcutaneous adipose tissue of severely burned patients, which would normally be discarded during surgical debridement. While previous studies found that severe burn injury alters the metabolic profile of subcutaneous adipocytes, the effect of severe burns on ASCs from burned victims is largely unknown. The purpose of this study is to compare the metabolic activity of ASCs from burned patients to those from non‐burned abdominoplasty patients by analyzing mitochondrial respiration and glycolysis. ASCs were isolated from adipose tissues from severely burned patients (BP, n=6) and abdominoplasty patients (AP, n=6). Cells were cultured to 80% confluence and harvested at at passage 2, 4, and 6 for analysis. Flow cytometry was used to determine ASC cell surface markers CD90, CD105, and CD73 and negative for CD34, CD11, CD19 and CD45. Mitochondrial abundance and reactive oxygen species (ROS) production were determined with MitoTracker Green and MitoSOX Red, respectively. JC‐10 Mitochondrial Membrane Potential Assays were used to determine mitochondrial membrane integrity. Mitochondrial respiration and glycolysis were analyzed in technical quadruplicates by a Seahorse XFe24 Analyzer. No significant differences were found in ASC surface markers, mitochondrial abundance or membrane potential among passages or between groups. A two‐way ANOVA showed that there was a significant effect (p<0.01) of passage number on mitochondrial ROS production. Mitochondrial respiratory states Basal, Leak (uncoupled), and Non‐mitochondrial respiration was significantly higher (p<0.05) at passage 6 than passage 2 for BP, but not AP. Additionally, Basal respiration was significantly higher at passage 6 in BP than AP (63.5±3.49 vs 45.7±4.26 pmol O2/min, p<0.001). Glycolysis significantly decreased (p<0.05) from passage 4 to 6 in AP but not BP ASCs. In summary, BP ASCs showed increased mitochondrial respiration with increasing passages and basal respiration was higher than AP ASCs patients at passage 6. While all ASCs retained their stem cell properties, they also showed increased ROS production through multiple passages which may cause oxidative stress that could limit the expansion of these cells ex vivo. The elevated mitochondrial activity from ASCs of burned patients may suggest these cells are more metabolically primed than those from abdominoplasty patients, but whether this thwarts their therapeutic potential requires additional investigation. Adipose‐derived stem cells from burned injured patients remain a potential viable source for burn wound therapies. While other studies have demonstrated efficacy in healing burn wounds, this study uncovers potential limitations in expansion of these cells ex vivo to use as coverage options for burned victims.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

392 Increased Mitochondrial Respiration and ROS Production From Adipose-Derived Stem Cells is Passage Dependent

Victims of severe burn injury (over 30% of the total body surface area) experience profound hypermetabolism which prolongs wound healing. Adipose-derived stem cells (ASCs) have been proposed as an attractive solution for treating burn wounds. Moreover, the subcutaneous adipose tissue of severely burned patients normally discarded during surgical debridement is a viable source of ASCs. While previous studies found that severe burn injury alters the metabolic profile of subcutaneous adipocyte, the effect of severe burns on ASCs is largely unknown. The purpose of this study is to compare the metabolic activity of ASCs from burned patients to those from abdominoplasty patients by analyzing mitochondrial respiration and characteristics. Isolated ASCs from severely burned patients (BP, n=6) and abdominoplasty patients (AP, n=6) were grown to 80% confluency, trypsinized, and harvested for analysis at passage 2, 4, and 6. Flow cytometry was used to determine ASC cell surface markers CD90, CD105, and CD73. Mitochondrial abundance and ROS production were determined with MitoTracker Green and MitoSOX Red, respectively. JC-10 Mitochondrial Membrane Potential Assays were used to determine mitochondrial membrane integrity. Mitochondrial respiration and glycolysis were analyzed in technical quadruplicates by a Seahorse XFe24 Analyzer. No significant differences were found in ASC surface markers, mitochondrial abundance or membrane potential among passages or between groups. A two-way ANOVA showed that there was a significant effect (p<0.01) of passages on mitochondrial ROS production. Mitochondrial respiratory states Routine, Leak (uncoupled), and Non-mitochondrial respiration was significantly higher (p<0.05) at passage 6 than passage 2 for BP, but not AP. Glycolysis significantly decreased (p<0.05) from passage 4 to 2 in AP ASCs. Stem cell positive staining and mitochondrial activity were not different between cells from burned or non-burned humans. BP ASCs showed increased mitochondrial respiration with increasing passages. All ASCs showed increased ROS production through multiple passages. Adipose-derived stem cells from burned injured patients remain an exciting potential source for therapeutic purposes. However, increased ROS production and mitochondrial leak may cause oxidative stress that could limit the expansion of these cells ex vivo.

Fat graft – the natural choice for reconstructive, regenerative and aesthetic surgery

AbstractThe search for appropriate filler, which can be used for aesthetic and reconstructive operations is currently one of challenges for plastic surgery. The application of absorbable and permanent artificial fillers may cause adverse events. Thus, autologous fat grafting can be a safe alternative. Moreover, fat tissue is rich in adipose-derived stem cells (ASC), which can be successfully used for regenerative procedures. The paper reviews reports on fat grafting procedures, which indicate risks and their possible prophylactic.Adipose tissue is a much more prolific source of ASCs than bone marrow. Basically, ASC are characterized by a spectrum of markers: CD11b-CD45-CD13+CD73+CD90+, which can be widened by CD36+CD-106-CD10+CD26+CD49d+CD49e+CD3-D49f -PODXL- to improve phenotyping. It is suggested to use at least two negative markers and two positive markers during the same phenotyping analysis. Fat transfer requires appropriate approach, planning and technique to make it clinically successful.Fat grafting fulfills the expectations for ideal injectable agent, which can be used for aesthetic and reconstructive surgery. To improve the survival of fat graft, careful decisions on donor site, local anesthetic administration, liposuction method, processing and placement methods need to be made. Moreover, fat is the source of adipose-derived stem cells which can be used for regenerative procedures. A proper transformation and identification of those cells is required to improve clinical effects.

Adipose-Derived Stem Cells in Aesthetic Surgery: A Mixed Methods Evaluation of the Current Clinical Trial, Intellectual Property, and Regulatory Landscape.

Adipose tissue, which can be readily harvested via a number of liposuction techniques, offers an easily accessible and abundant source of adipose-derived stem cells (ASCs). Consequently, ASCs have become an increasingly popular reconstructive option and a novel means of aesthetic soft tissue augmentation. This paper examines recent advances in the aesthetic surgery field, extending beyond traditional review formats to incorporate a comprehensive analysis of current clinical trials, adoption status, and the commercialization pathway. Keyword searches were carried out on clinical trial databases to search for trials using ASCs for aesthetic indications. An intellectual property landscape was created using commercial software (Thomson Reuters Thomson Innovation, New York, NY). Analysis of who is claiming what in respect of ASC use in aesthetic surgery for commercial purposes was analyzed by reviewing the patent landscape in relation to these techniques. Key international regulatory guidelines were also summarized. Completed clinical trials lacked robust controls, employed small sample sizes, and lacked long-term follow-up data. Ongoing clinical trials still do not address such issues. In recent years, claims to intellectual property ownership have increased in the "aesthetic stem cell" domain, reflecting commercial interest in the area. However, significant translational barriers remain including regulatory challenges and ethical considerations. Further rigorous randomized controlled trials are required to delineate long-term clinical efficacy and safety. Providers should consider the introduction of patient reported outcome metrics to facilitate clinical adoption. Robust regulatory and ethical policies concerning stem cells and aesthetic surgery should be devised to discourage further growth of "stem cell tourism."

Tissue Source and Cell Expansion Condition Influence Phenotypic Changes of Adipose-Derived Stem Cells.

Stem cells derived from the subcutaneous adipose tissue of debrided burned skin represent an appealing source of adipose-derived stem cells (ASCs) for regenerative medicine. Traditional tissue culture uses fetal bovine serum (FBS), which complicates utilization of ASCs in human medicine. Human platelet lysate (hPL) is one potential xeno-free, alternative supplement for use in ASC culture. In this study, adipogenic and osteogenic differentiation in media supplemented with 10% FBS or 10% hPL was compared in human ASCs derived from abdominoplasty (HAP) or from adipose associated with debrided burned skin (BH). Most (95–99%) cells cultured in FBS were stained positive for CD73, CD90, CD105, and CD142. FBS supplementation was associated with increased triglyceride content and expression of adipogenic genes. Culture in hPL significantly decreased surface staining of CD105 by 31% and 48% and CD142 by 27% and 35% in HAP and BH, respectively (p < 0.05). Culture of BH-ASCs in hPL also increased expression of markers of osteogenesis and increased ALP activity. These data indicate that application of ASCs for wound healing may be influenced by ASC source as well as culture conditions used to expand them. As such, these factors must be taken into consideration before ASCs are used for regenerative purposes.

Editorial: Cell-Based Therapies for Diabetic Microvascular Complications.

Editorial: Cell-Based Therapies for Diabetic Microvascular Complications.

Adipose-derived stems cells and their role in human cancer development, growth, progression, and metastasis: a systematic review.

Obesity is a well recognized risk factor for several types of cancers, many of which occur solely or disproportionately in women. Adipose tissue is a rich source of adipose-derived stem cells (ASC), which have received attention for their role in cancer behavior. The purpose of this systematic review is to present the existing literature on the role of ASCs in the growth, development, progression, and metastasis of cancer, with an emphasis on malignancies that primarily affect women. To accomplish this goal, the bibliographic database PubMed was systematically searched for articles published between 2001 and 2014 that address ASCs' relationship to human cancer. Thirty-seven articles on ASCs' role in human cancer were reviewed. Literature suggests that ASCs exhibit cancer-promoting properties, influence/are influenced by the tumor microenvironment, promote angiogenesis, and may be associated with pathogenic processes through a variety of mechanisms, such as playing a role in hypoxic tumor microenvironment. ASCs appear to be important contributors to tumor behavior, but research in areas specific to women's cancers, specifically endometrial cancer, is scarce. Also, because obesity continues to be a major health concern, it is important to continue research in this area to improve understanding of the impact adiposity has on cancer incidence.

Comparison of Markers and Functional Attributes of Human Adipose-Derived Stem Cells and Dedifferentiated Adipocyte Cells from Subcutaneous Fat of an Obese Diabetic Donor

Objective: Adipose tissue is a robust source of adipose-derived stem cells (ADSCs) that may be able to provide secreted factors that promote the ability of wounded tissue to heal. However, adipocytes also have the potential to dedifferentiate in culture to cells with stem cell-like properties that may improve their behavior and functionality for certain applications. Approach: ADSCs are adult mesenchymal stem cells that are cultured from the stromal vascular fraction of adipose tissue. However, adipocytes are capable of dedifferentiating into cells with stem cell properties. In this case study, we compare ADSC and dedifferentiated fat (DFAT) cells from the same patient and fat depot for mesenchymal cell markers, embryonic stem cell markers, ability to differentiate to adipocytes and osteoblasts, senescence and telomerase levels, and ability of conditioned media (CM) to stimulate migration of human dermal fibroblasts (HDFs). Innovation and Conclusions: ADSCs and DFAT cells displayed identical levels of CD90, CD44, CD105, and were CD34- and CD45-negative. They also expressed similar levels of Oct4, BMI1, KLF4, and SALL4. DFAT cells, however, showed higher efficiency in adipogenic and osteogenic capacity. Telomerase levels of DFAT cells were double those of ADSCs, and senescence declined in DFAT cells. CM from both cell types altered the migration of fibroblasts. Despite reports of ADSCs from a number of human depots, there have been no comparisons of the ability of dedifferentiated DFAT cells from the same donor and depot to differentiate or modulate migration of HDFs. Since ADSCs were from an obese diabetic donor, reprogramming of DFAT cells may help improve a patient's cells for regenerative medicine applications.

Expansion of Semi-Automatic Processed Human Adipose-Derived Stem Cells in Medium Supplemented with Autologous Serum and Antioxidants

Objective: Excised fat tissue (EFT) is an excellent source of adipose-derived stem cells (ADSCs) for autologous cell therapy. However, the current technique for excising fat tissues with conventional surgical scissors and using culture medium containing fetal bovine serum (FBS) may not comply with Good Tissue Practice (GTP) guidelines. We thus developed a semi-automated process complying with the abovementioned guideline to manufacture clinicalgrade ADSCs from small volumes of EFT. Methods: A semi-automated process was developed by using a disposable disperser tube operated on a Tube Disperser Workstation to obtain stromal vascular fraction (SVF) from EFT. A culture medium supplemented with human autologous serum (HAS) and mesenchymal stem cell culture adjuvant (MCA) consisted of basic fibroblast growth factor (bFGF/FGF-2) and antioxidants, was formulated to expand ADSCs from SVF in vitro. Results: The semi-automated process can markedly enhance cell yields and reduce the operation time for mincing tissues from an hour down to a few minutes. The growth of ADSCs was slower in the 10% HAS medium than that in medium supplemented with FBS, but their growth in the 10% HAS medium containing MCA (10% HAS+MCA) was superior to both media. The 10% HAS+MCA medium also promoted adipogenesis, osteogenesis, chondrogenesis and increased the expression of CD44, CD73 and the stemness genes in the ADSCs. Conclusion: Our novel semi-automated process provides an efficient isolation of SVF, and the 10% HAS+MCA medium accelerates the growth of ADSCs from small amounts of EFT without losing their differentiation ability.