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Adipose stem cell

Marie Rogne, Dinh-Toi Chu, Thomas M Küntziger, Maria-Niki Mylonakou, Philippe Collas, Kjetil Tasken
Optic atrophy 1 (OPA1) is the A-kinase anchoring protein (AKAP) targeting the pool of PKA responsible for perilipin 1 phosphorylation, a gatekeeper for lipolysis. However, the involvement of OPA1-bound PKA in the downstream regulation of lipolysis is unknown. Here, we show upregulation and relocation of OPA1 from mitochondria to lipid droplets during adipocytic differentiation of human adipose stem cells (hASCs). We employed various biochemical and immunological approaches to demonstrate that OPA1-bound PKA phosphorylates perilipin 1 at S522 and S497 upon lipolytic stimulation...
April 24, 2018: Molecular Biology of the Cell
Shifeng Yan, Pengfei Xia, Shenghua Xu, Kunxi Zhang, Guifei Li, Lei Cui, Jingbo Yin
Porous microcarriers have aroused increasing attention recently, which can create a protected environment for sufficient cell seeding density, facilitate oxygen and nutrient transfer, and well support the cell attachment and growth. In this study, porous microcarriers fabricated from the strontium-substituted hydroxyapatite-graft-poly(γ-benzyl-L-glutamate) (Sr10-HA-g-PBLG) hybrid nanocomposite were developed. The surface grating of PBLG, the micromorphology and element distribution, mechanical strength, in vitro degradation and Sr2+ ion release of the obtained Sr10-HA-g-PBLG porous microcarriers were investigated, respectively...
April 24, 2018: ACS Applied Materials & Interfaces
Vajiheh Neshati, Samaneh Mollazadeh, Bibi Sedigheh Fazly Bazzaz, Mehrdad Iranshahi, Majid Mojarrad, Hojjat Naderi-Meshkin, Mohammad Amin Kerachian
Stem cell therapy is considered as a promising treatment for cardiovascular diseases. Adipose-derived mesenchymal stem cells (ADMSCs) have the ability to undergo cardiomyogenesis. Medicinal plants are effective and safe candidates for cell differentiation. Therefore, the aim of our study was to investigate cardiogenic effects of characterized (HPLC-UV) G. urbanum extracts on ADMSCs of adipose tissue. The methanolic extracts of the root and aerial parts of G. urbanum were obtained and MTT assay was used for studying their cytotoxic effects...
April 24, 2018: Biochemistry and Cell Biology, Biochimie et Biologie Cellulaire
Satoshi Tajima, Morikuni Tobita, Hiroshi Mizuno
Mesenchymal stem cells (MSCs) have the potential to directly differentiate into osteogenic cells and efficiently regenerate bone tissue. Adipose-derived stem cells (ASCs) have the potential to differentiate into an osteogenic lineage, too. In addition, ASCs can be readily harvested in large numbers with low donor-site morbidity. Meanwhile, recent reports have demonstrated that platelet-rich plasma (PRP) contains a variety of growth factors and may be a powerful biological autologous cocktail of growth factors for tissue engineering...
2018: Methods in Molecular Biology
Shahensha Shaik, Ram Devireddy
The development of simple but effective storage protocols for adult stem cells will greatly enhance their use and utility in tissue-engineering applications. Cryopreservation has shown to be most promising but is a fairly complex process, necessitating the use of chemicals called cryoprotective agents (CPAs), freezing equipment, and obviously, storage in liquid nitrogen. The purpose of this chapter is to present a general overview of cryopreservation storage techniques and the optimal protocols/results obtained in our laboratory for long-term storage of adult stem cells using freezing storage...
2018: Methods in Molecular Biology
Rachel C Nordberg, Josie C Bodle, Elizabeth G Loboa
It is critical that human adipose stem cell (hASC) tissue-engineering therapies possess appropriate mechanical properties in order to restore function of the load bearing tissues of the musculoskeletal system. In an effort to elucidate the hASC response to mechanical stimulation and develop mechanically robust tissue engineered constructs, recent research has utilized a variety of mechanical loading paradigms including cyclic tensile strain, cyclic hydrostatic pressure, and mechanical unloading in simulated microgravity...
2018: Methods in Molecular Biology
Francesco Egro, Kacey G Marra
Autologous fat transplantation has revolutionized soft tissue reconstruction, but conventional methods remain unpredictable as graft resorption rates are high due to lack of vascularization. The advent of adipose-derived stem cells (ASCs) has led to improvement of fat grafting outcomes, in part to their ability to undergo facile differentiation into adipose tissue, their angiogenic properties, and their ability to express and secrete multiple growth factors. This chapter discusses the isolation and characterization of human ASCs, its expansion in vitro, and relevant in vivo models for adipose tissue engineering...
2018: Methods in Molecular Biology
Jie Li, Hui Li, Weidong Tian
Adipose tissue provides valuable cell source for tissue engineering, regenerative medicine, and adipose tissue biology studies. The most widely used adipose-derived stromal/stem cells (ASCs) isolation protocol involves enzymatic digestion with collagenase. However, the yield of the method is poor even impossible to collect enough stromal vascular fraction (SVF) for expansion when the sample size is small, for instance only new born mice is available for cell culture. Here, we describe an efficient protocol for the isolation and expansion of ASCs using explant culture...
2018: Methods in Molecular Biology
Jie Li, J Lowry Curley, Z Elizabeth Floyd, Xiying Wu, Yuan Di C Halvorsen, Jeffrey M Gimble
Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Here, we describe methods from our own laboratory and the literature for the isolation and expansion of adipose-derived stem cells (ASCs). We present a large scale procedure suitable for processing >100 mL volumes of lipoaspirate tissue specimens by collagenase digestion and a related procedure suitable for processing adipose tissue aspirates without digestion...
2018: Methods in Molecular Biology
Hubert Tseng, Alexes C Daquinag, Glauco R Souza, Mikhail G Kolonin
White adipose tissue (WAT) has attracted interest for tissue engineering and cell-based therapies as an abundant source of adipose stem/stromal cells (ASC). However, technical challenges in WAT cell culture have limited its applications in regenerative medicine. Traditional two-dimensional (2D) cell culture models, which are essentially monolayers of cells on glass or plastic substrates, inadequately represent tissue architecture, biochemical concentration gradients, substrate stiffness, and most importantly for WAT research, cell phenotypic heterogeneity...
2018: Methods in Molecular Biology
Gail Kilroy, Marilyn Dietrich, Xiying Wu, Jeffrey M Gimble, Z Elizabeth Floyd
Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Adipose tissue stromal vascular cells or primary adipocytes derived from murine adipose tissue and grown in culture are essential tools for studying the mechanisms underlying adipocyte development and function. Herein, we describe methods for the isolation, expansion, and long-term storage of murine adipose-derived stromal/stem cells along with protocols for inducing adipogenesis in this cell population or isolating the adipose stromal vascular fraction cells for flow cytometric analysis...
2018: Methods in Molecular Biology
Pinar Yilgor Huri, Justin Morrissette-McAlmon, Warren L Grayson
Adipose-derived stem/stromal cells (ASCs) constitute a very promising source for cell therapy and tissue engineering approaches as they can be easily obtained in large quantities with comparatively minimal patient discomfort. Moreover, ASCs have multilineage differentiation capacity. Among these, differentiation capacity along the myogenic lineage is of particular interest since myogenic precursors are scarce and obtaining a large number of cells from skeletal muscle biopsies is difficult. Here, we describe a method to effectively induce ASC myogenesis through the combination of biochemical (cocktail including 5-azacytidine and horse serum) and biophysical (dynamic culture via uniaxial cyclic strain) stimulation...
2018: Methods in Molecular Biology
Pietro Gentile, Valerio Cervelli
Cell-based therapy and regenerative medicine offer a paradigm shift in regard to various diseases causing loss of substance or volume and tissue or organ damage. Recently, many authors have focused their attention on mesenchymal stem cells for their capacity to differentiate into many cell lineages. The most widely studied types are bone marrow mesenchymal stem cells and adipose derived stem cells (ADSCs), which display similar results. Based on the literature, we believe that the ADSCs offer advantages because of lower morbidity during the harvesting procedure...
2018: Methods in Molecular Biology
Pascal Morissette Martin, Arthi Shridhar, Claire Yu, Cody Brown, Lauren E Flynn
Surgically discarded adipose tissue is not only an abundant source of multipotent adipose-derived stem/stromal cells (ASCs) but can also be decellularized to obtain a biomimetic microenvironment for tissue engineering applications. The decellularization methods involve processing excised fat through a series of chemical, mechanical, and enzymatic treatment stages designed to extract cells, cellular components, and lipid from the tissues. This process yields a complex 3D bioscaffold enriched in collagens that mimics the biochemical and biomechanical properties of the native extracellular matrix (ECM)...
2018: Methods in Molecular Biology
Christina L Rettinger, Kai P Leung, Rodney K Chan
Adipose-derived stem cells are capable of self-renewal and differentiation along multiple cell lineages, and have potential applications in a wide range of therapies. ASCs are commonly cultured as monolayers on tissue culture plastic, but there are indications that they may lose their cell-specific properties with time in vitro. There has been a growing interest in culturing adherent cells using three-dimensional techniques based on the understanding that growing cells on plastic surfaces cannot truly recapitulate 3D in vivo conditions...
2018: Methods in Molecular Biology
Joel A Aronowitz, Ryan A Lockhart, Cloe S Hakakian
There is increasing interest in the clinical applications of adipose-derived stem cells (ASCs) and the stromal vascular fraction (SVF) based on promising preclinical data. As adipose-derived therapeutics begin to translate into the clinical setting, it is important to maintain patient safety as well as uniformity in technique. Here, we describe a method for isolation of stromal vascular fraction cells in a clinically relevant time frame. Analytical laboratory techniques are mentioned, but respective protocols are not provided here...
2018: Methods in Molecular Biology
Annie C Bowles, Alan Tucker, Bruce A Bunnell
Evidence from preclinical research and clinical trials demonstrates the use of the stromal vascular fraction (SVF) as therapy for numerous indications. These results demonstrate that autologous SVF is not only safe and effective but provides robust anti-inflammatory, immunomodulatory, and reparative effects in vivo. The potency of the SVF is attributed to the cellular composition which includes adipose-derived stem cells (ASCs), adipocytes, endothelial cells, and various immune cells. As the name would suggest, these SVF cells are derived from the stromal compartment of adipose, or fat...
2018: Methods in Molecular Biology
Walter Baumgartner, Isabelle Schneider, Samuel C Hess, Wendelin J Stark, Sonja Märsmann, Marzia Brunelli, Maurizio Calcagni, Paolo Cinelli, Johanna Buschmann
OBJECTIVE: Chemical supplementation of culture media to induce differentiation of adult stem cells seeded on a scaffold may mask other differentiation triggers such as scaffold stiffness, chemical composition or mechanical stimulation. However, stem cells can be differentiated towards osteoblasts without any supplementation given an appropriate osteogenic scaffold and an adequate mechanical stimulation. MATERIALS AND METHODS: Electrospun meshes of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/aCaP) in a weight ratio of 60:40 were seeded with human adipose-derived stem cells (ASCs) and cultured in DMEM...
April 5, 2018: Journal of the Mechanical Behavior of Biomedical Materials
Anni Sorkio, Lothar Koch, Laura Koivusalo, Andrea Deiwick, Susanna Miettinen, Boris Chichkov, Heli Skottman
There is a high demand for developing methods to produce more native-like 3D corneal structures. In the present study, we produced 3D cornea-mimicking tissues using human stem cells and laser-assisted bioprinting (LaBP). Human embryonic stem cell derived limbal epithelial stem cells (hESC-LESC) were used as a cell source for printing epithelium-mimicking structures, whereas human adipose tissue derived stem cells (hASCs) were used for constructing layered stroma-mimicking structures. The development and optimization of functional bioinks was a crucial step towards successful bioprinting of 3D corneal structures...
April 16, 2018: Biomaterials
S Wan, X Fu, Y Ji, M Li, X Shi, Y Wang
Adipose-derived mesenchymal stem cells (ASCs) represent an excellent candidate for tissue engineering due to their multipotency and paracrine functions. Physical cues provided by the scaffolds have been shown to direct a variety of cellular behaviors of ASCs; however, their effects on the paracrine functions of ASCs, especially immunomodulatory functions, remain largely unexplored. Thus, PLLA electrospun fibrous scaffolds, with fibers oriented either randomly or aligned, were used as a model to investigate the effects of fiber orientation on the immunomodulatory paracrine secretion of ASCs...
April 16, 2018: Biomaterials
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