|  |  |  | | | | | TE-RegenMed-StemCell feed | | | | | | | | | | | | | | | | | | | Stroke patients take part in "milestone" UK trial of stem cell therapy. BMJ. 2010;341:c6574 Authors: Wise J PMID: 21081623 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Can intracoronary stem cell injection permanently improve cardiac function after myocardial infarction? Interact Cardiovasc Thorac Surg. 2010 Nov 16; Authors: Abbasi M, Javan H, Alizadeh B, Afzalnia S A best evidence topic in cardiac surgery was written according to a structured protocol. The question addressed was 'Can intracoronary stem cell injection permanently improve cardiac function after myocardial infarction?'. Altogether 314 papers were found using the reported search, of which five represented the best evidence to answer the clinical question. The authors, journal, date and country of publication, patient group studied, study type, relevant outcomes and results of these papers are tabulated. We conclude that stem cells may have controversial effects on cardiac function in long-term follow-up of more than two years as they improved the left ventricular ejection fraction and end systolic volume index just in two studies in which none of them utilized cardiac magnetic resonance imaging (MRI), as the most reliable method, to quantify cardiac function. However, all remaining three trials which reported negative results used cardiac MRI for assessment of cardiac indexes which may be more precise and accurate than echocardiographic assessments. So the reliability of the positive trials is lower than negative resulted trials in terms of cardiac function assessment method. Stem cell therapy almost always offered short-term benefits over the best medical treatment, but the long-term benefits are still a matter of debate. Keywords: Myocardial infarction; Stem cells; Ventricular function. PMID: 21081555 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Cell Sheet Technology for Tissue Engineering: The Self-Assembly Approach Using Adipose-Derived Stromal Cells. Methods Mol Biol. 2011;702:429-441 Authors: Labbé B, Marceau-Fortier G, Fradette J In the past years, adipose tissue has spurred a wide interest, not only as a source of adult multipotent stem cells but also as a highly eligible tissue for reconstructive surgery procedures. Tissue engineering is one field of regenerative medicine progressing at great strides in part due to its important use of adipose-derived stem/stromal cells (ASCs). The development of diversified technologies combining ASCs with various biomaterials has lead to the reconstruction of numerous types of tissue-engineered substitutes such as bone, cartilage, and adipose tissues from rodent, porcine, or human ASCs. We have recently achieved the reconstruction of connective and adipose tissues composed entirely of cultured human ASCs and their secreted endogenous extracellular matrix components by a methodology known as the self-assembly approach of tissue engineering. The latter is based on the stimulation of ASCs to secrete and assemble matrix components in culture, leading to the production of cell sheets that can be manipulated and further assembled into thicker multilayer tissues. In this chapter, protocols to generate both reconstructed connective and adipocyte-containing tissues using the self-assembly approach are described in detail. The methods include amplification and cell banking of human ASCs, as well as culture protocols for the production of individual stromal and adipose sheets, which are the building blocks for the reconstruction of multilayered human connective and adipose tissues, respectively. PMID: 21082420 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Soft Tissue Reconstruction. Methods Mol Biol. 2011;702:395-400 Authors: Rubin JP, Marra KG The potential of adipose-derived stem cells (ASCs) in clinical applications of soft tissue regeneration is immense. This chapter discusses the isolation and characterization of human ASCs, expansion in vitro, and relevant in vivo models for adipose tissue engineering. PMID: 21082417 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | A First Approach for the Production of Human Adipose Tissue-Derived Stromal Cells for Therapeutic Use. Methods Mol Biol. 2011;702:331-343 Authors: Bourin P, Peyrafitte JA, Fleury-Cappellesso S Adipose tissue-derived stromal cells (ASCs) are promising tools for the new therapeutic field of regenerative medicine. Many research teams are intent on producing these cells for therapeutic purposes. The cell production must follow strict rules for safety and for constant quality of the cell product to ensure a reliable effect in patients. These rules are grouped under the generic term Good Manufacturing Practices. In this chapter, we describe the general concepts of ASC production for therapeutic use, explaining new terms such as traceability and qualification. We also introduce general requirements for the installation, equipment, material, and staff for the cell production. Then, we outline a general strategy for building a cell culture process. Finally, as an example, we describe the use of CellStack™ chambers and specific tube sets that allow for producing cells beginning with the stromal vascular fraction under near-closed conditions. PMID: 21082413 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Adipose Stem Cell Differentiation into Smooth Muscle Cells. Methods Mol Biol. 2011;702:261-268 Authors: Marra KG, Brayfield CA, Rubin JP The differentiation of adipose-derived stem cells (ASCs) into functional smooth muscle cells has received limited investigation. Various methodologies for both in vitro and in vivo differentiation is described. In vitro differentiation is obtained by either chemical or mechanical stimulation, and is determined by expression of smooth muscle cell markers. In vivo differentiation studies include animal models of cardiovascular disease and one study with urinary bladder reconstruction. The ease of obtaining an abundant number of ASCs render this cell population useful for potential vascular therapies that require autologous smooth muscle cells. PMID: 21082408 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Osteogenic Differentiation Strategies for Adipose-Derived Mesenchymal Stem Cells. Methods Mol Biol. 2011;702:233-248 Authors: Kroeze RJ, Knippenberg M, Helder MN Adipose stem cell preparations, either obtained as a freshly isolated so-called stromal vascular fraction (SVF) or as cells cultured to homogeneity and then referred to as adipose stem cells (ASCs), have found widespread use in a broad variety of studies on tissue engineering and regenerative medicine applications, including bone repair.For newcomers within the field, but also for established research laboratories having up to 10 years of expertise in this research area, it may be convenient to strive for, and use consensus protocols (1) for studying the osteogenic differentiation potential of ASC preparations in vitro, and (2) for osteogenic induction regimes for in vivo implementation. To assist in achieving this goal, this chapter describes various step-by-step osteogenic differentiation protocols for adipose-derived stem cell populations (SVF as well as ASCs) currently applied within our laboratory, with particular emphasis on protocols aimed at intra-operative use. The protocols describe the use of inducing compounds, including the bone morphogenetic proteins (BMPs), 1,25-dihydroxyvitamin-D3, and polyamines, as well as methods and parameters for evaluating the level of differentiation achieved.We would appreciate receiving feedback on the protocols described; this will facilitate the development of consensus protocols, which in turn will allow better comparison of data sets generated by different research groups. This continuing standardization, which might be reported on at international meetings like those of IFATS ( http://www.IFATS.org ), might be of benefit for the whole ASC research community. PMID: 21082406 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Neural Differentiation of Human Adipose Tissue-Derived Stem Cells. Methods Mol Biol. 2011;702:219-231 Authors: Yu JM, Bunnell BA, Kang SK While adult stem cells can be induced to transdifferentiate into multiple lineages of cells or tissues, their plasticity and utility for human therapy remains controversial. In this chapter, we describe methods for the transdifferentiation of human adipose tissue-derived stem cells (ASCs) along neural lineages using in vitro and in vivo systems. The in vitro neural differentiation of ASCs has been reported by several groups using serum-free cytokine induction, butylated hydroxyanisole (BHA) chemical induction, and neurosphere formation in combination with the cytokines, such as brain-derived neurotrophic factor (BDNF) and basic fibroblast growth factor (bFGF). For in vivo neurogenic induction, ASCs are treated with BDNF and bFGF to form neurospheres in vitro and then delivered directly to the brain. In this chapter, several detailed protocols for the effective neurogenic induction of ASCs in vitro and in vivo are described. The protocols described herein can be applied to further molecular and mechanistic studies of neurogenic induction and differentiation of ASCs. In addition, these methods can be useful for differentiating ASCs for therapeutic intervention in central nervous system disorders. PMID: 21082405 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Characterization of Human Adipose-Derived Stem Cells Using Flow Cytometry. Methods Mol Biol. 2011;702:121-131 Authors: Tucker HA, Bunnell BA One of the hallmark characteristics of human adipose-derived stem cells (hASCs) is their ability to differentiate into cells of mesenchymal lineages. It is also becoming apparent that ASCs can mediate a therapeutic benefit through cytokine, paracrine-driven mechanisms influencing apoptosis, angiogenesis, and potent anti-inflammatory responses. Although there is still no clear consensus on the antigen expression pattern that will define hASCs, a protocol is also presented for the flow cytometric analysis utilizing a series of antibody panels. The analysis of these surface epitope patterns can aide in the isolation and characterization of hASCs. Moreover, using this standardized antibody panel, direct comparisons can be made between ASCs isolated from various tissue sources, which will benefit the field by providing uniformity to the comparison process. PMID: 21082399 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | In Vitro Adult Canine Adipose Tissue-Derived Stromal Cell Growth Characteristics. Methods Mol Biol. 2011;702:47-60 Authors: Spencer ND, Lopez MJ Stromal cells are undifferentiated cells found in embryonic and adult tissues. Adult mesenchymal stromal cells (MSCs) possess the properties of self renewal, long-term viability, multipotentiality, and immune privilege, which make them attractive candidates for regenerative medicine applications. In order to develop targeted adult stromal cell therapies for diseased and injured tissues in animals and humans, it is essential to have large-animal models. The dog represents not only a patient population, but is also a valuable experimental model. The dog has contributed significantly to the understanding of various human diseases such as genetic and musculoskeletal disorders. In order to optimize the use of stromal cell therapy in the dog as a patient or disease model, a comprehensive characterization of the cells is required. PMID: 21082394 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Isolation and Culture of Rhesus Adipose-Derived Stem Cells. Methods Mol Biol. 2011;702:3-16 Authors: Gagliardi C, Bunnell BA Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Rhesus monkeys are physiologically and phylogenetically similar to humans, and they and their cells are valuable for biomedical research and evaluation of preclinical therapies. Here, we describe methods for the isolation, culture, and differentiation of rhesus adipose-derived stem cells (rASCs). PMID: 21082390 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | The therapeutic potential of engineered human neovessels for cell-based gene therapy. Expert Opin Biol Ther. 2010 Nov 17; Authors: Alvarez-Vallina L, Sanz L Importance of the field: Several works have shown the feasibility of engineering functional blood vessels in vivo using human endothelial cells and mural cells. In this context, the genetic modification of endothelial cells would ensure the secretion of a therapeutic protein into the systemic circulation for a prolonged period of time. Areas covered in this review: We discuss the different strategies aimed at the formation of long-lasting neovessels in vivo, using human endothelial and mural cells. The main focus is the potential of these constructs in gene therapy strategies for the in vivo production of therapeutic proteins. What the reader will gain: The reader will have an outline of the different types of cells that have been used for microvessel engineering in vivo, as well as scaffolds employed to seed these cells. We provide a critical review of their advantages and drawbacks, along with examples of their potential in cell-based gene therapy strategies. Take home message: There is a real potential for neovessels derived from human endothelial and mural cells to be incorporated in clinical interventions, either as a cell-based gene therapy to produce a therapeutic protein or as a component of engineered tissue constructs in regenerative medicine. PMID: 21080857 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Comparison of beneficial effects of undifferentiated cultured bone marrow stromal cells and omental adipose-derived nucleated cell fractions on sciatic nerve regeneration. Muscle Nerve. 2010 Nov 16; Authors: Mohammadi R, Azizi S, Delirezh N, Hobbenaghi R, Amini K Adipose tissue is a good source for isolation of cells with stem-cell-like properties. The effects of undifferentiated cultured bone marrow stromal cells (BMSCs) and omental adipose-derived nucleated cells (OADNCs) on peripheral nerve regeneration were compared in a rat nerve regeneration model. A 10-mm sciatic nerve defect was bridged using a vein graft. In one group, the vein was filled with BMSCs and in the other group with OADNCs. Functional study, morphometric indices, and immunohistochemistry indicated there was no significant difference (P > 0.05) between groups in recovery of regenerated axons at 4, 8, and 12 weeks after surgery. OADNCs enhanced regeneration similar to undifferentiated BMSCs. These observations suggest OADNCs represent an effective and cost-saving cell population due to the shortened time interval from tissue collection to cell injection as well as procedural simplicity. This approach is clinically translatable toward new methods for enhanced peripheral nerve repair without the limitations of BMSC. Muscle Nerve, 2010. PMID: 21082699 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Use of Adipose-Derived Stem Cells in High-Throughput Screening to Identify Modulators of Lipogenesis. Methods Mol Biol. 2011;702:359-368 Authors: Lea-Currie YR, Duffin DJ, Buehrer BM Drug discovery efforts have an increasing focus on functional cell-based screening to identify compounds that modulate targets presented in a relevant format. Historically, immortalized cell lines have been used in primary and secondary screens due to their ease of manipulation, transformation, and propagation. However, more researchers are using primary cells that present their drug targets in their natural context. Human primary cell isolation and propagation procedures have become efficient enough to provide these cells in the necessary scale for early stage drug discovery. Adult human stem cells provide an opportunity for investigating multiple pathways of differentiation, development, regeneration, and toxicity using a single cell source and type. Adipose-derived stem cells (ASCs) are an attractive adult human primary stem cell for drug discovery due their abundance in adipose tissue, ease of isolation, and propagation in culture. They can be expanded in high numbers and retain their unique properties to differentiate into multiple lineages. In this chapter, we describe a protocol to identify modulators of human ASC lipogenesis following partial differentiation to adipocytes. PMID: 21082415 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Adipose Stem Cell Differentiation into Smooth Muscle Cells. Methods Mol Biol. 2011;702:261-268 Authors: Marra KG, Brayfield CA, Rubin JP The differentiation of adipose-derived stem cells (ASCs) into functional smooth muscle cells has received limited investigation. Various methodologies for both in vitro and in vivo differentiation is described. In vitro differentiation is obtained by either chemical or mechanical stimulation, and is determined by expression of smooth muscle cell markers. In vivo differentiation studies include animal models of cardiovascular disease and one study with urinary bladder reconstruction. The ease of obtaining an abundant number of ASCs render this cell population useful for potential vascular therapies that require autologous smooth muscle cells. PMID: 21082408 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Osteogenic Differentiation Strategies for Adipose-Derived Mesenchymal Stem Cells. Methods Mol Biol. 2011;702:233-248 Authors: Kroeze RJ, Knippenberg M, Helder MN Adipose stem cell preparations, either obtained as a freshly isolated so-called stromal vascular fraction (SVF) or as cells cultured to homogeneity and then referred to as adipose stem cells (ASCs), have found widespread use in a broad variety of studies on tissue engineering and regenerative medicine applications, including bone repair.For newcomers within the field, but also for established research laboratories having up to 10 years of expertise in this research area, it may be convenient to strive for, and use consensus protocols (1) for studying the osteogenic differentiation potential of ASC preparations in vitro, and (2) for osteogenic induction regimes for in vivo implementation. To assist in achieving this goal, this chapter describes various step-by-step osteogenic differentiation protocols for adipose-derived stem cell populations (SVF as well as ASCs) currently applied within our laboratory, with particular emphasis on protocols aimed at intra-operative use. The protocols describe the use of inducing compounds, including the bone morphogenetic proteins (BMPs), 1,25-dihydroxyvitamin-D3, and polyamines, as well as methods and parameters for evaluating the level of differentiation achieved.We would appreciate receiving feedback on the protocols described; this will facilitate the development of consensus protocols, which in turn will allow better comparison of data sets generated by different research groups. This continuing standardization, which might be reported on at international meetings like those of IFATS ( http://www.IFATS.org ), might be of benefit for the whole ASC research community. PMID: 21082406 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Adipogenic Differentiation of Adipose-Derived Stem Cells. Methods Mol Biol. 2011;702:193-200 Authors: Yu G, Floyd ZE, Wu X, Hebert T, Halvorsen YD, Buehrer BM, Gimble JM The primary physiological function of adipose-derived stem cells (ASCs) is to differentiate into adipose tissue. It is now possible to isolate, expand, and cryopreserve ASC from adipose depots of many animal species. These ASC can be induced to undergo adipogenic differentiation in vitro by exposure to a cocktail of chemical agents or inductive growth factors. The current chapter describes methods to induce adipogenesis and to quantify this differentiation process in vitro. PMID: 21082403 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Characterization of Human Adipose-Derived Stem Cells Using Flow Cytometry. Methods Mol Biol. 2011;702:121-131 Authors: Tucker HA, Bunnell BA One of the hallmark characteristics of human adipose-derived stem cells (hASCs) is their ability to differentiate into cells of mesenchymal lineages. It is also becoming apparent that ASCs can mediate a therapeutic benefit through cytokine, paracrine-driven mechanisms influencing apoptosis, angiogenesis, and potent anti-inflammatory responses. Although there is still no clear consensus on the antigen expression pattern that will define hASCs, a protocol is also presented for the flow cytometric analysis utilizing a series of antibody panels. The analysis of these surface epitope patterns can aide in the isolation and characterization of hASCs. Moreover, using this standardized antibody panel, direct comparisons can be made between ASCs isolated from various tissue sources, which will benefit the field by providing uniformity to the comparison process. PMID: 21082399 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Methods for the Purification and Characterization of Human Adipose-Derived Stem Cells. Methods Mol Biol. 2011;702:109-120 Authors: Gronthos S, Zannettino AC Peripheral adipose tissue contains a population of clonogenic precursor cells referred to as adipose-derived stem cells (ASC) that retain the capacity to differentiate into multiple cell types including osteoblasts, adipocytes, chondrocytes, myocytes, and neuronal cells following ex vivo expansion. Recent studies have demonstrated that ASC are most likely derived from a perivascular niche within highly vascularised fat tissue, analogous to different mesenchymal cell populations identified in other tissues throughout the body. The following chapter describes techniques to prospectively isolate clonogenic ASC from adult human adipose tissue using antibodies directed against perivascular markers and methods to immunophenotypically characterize their ex vivo expanded progeny. PMID: 21082398 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Isolation and Culture of Porcine Adipose Tissue-Derived Somatic Stem Cells. Methods Mol Biol. 2011;702:77-86 Authors: Williams KJ, Godke RA, Bondioli KR Adipose tissue-derived stem cells (ASCs) have been described for a number of laboratory animals and humans. Improved culture conditions and cellular characteristics of ASCs have been identified. ASCs can self-renew and differentiate into multiple tissue lineages. Further characterization of ASCs in this manner could enhance the isolation and purification of a population of mesenchymal stem cells (MSCs) from easily obtainable adipose tissue. These stem cell populations from domestic animals, which make attractive models for transplantation studies, will be valuable for the evaluation of their efficacy in tissue regeneration applications in the future. These cells may also represent a population more easily reprogrammable during somatic cell nuclear transfer and thus expedite the development of transgenic animals for models and production of valuable pharmaceutical proteins. PMID: 21082396 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Isolation of Murine Adipose-Derived Stem Cells. Methods Mol Biol. 2011;702:29-36 Authors: Yu G, Wu X, Kilroy G, Halvorsen YD, Gimble JM, Floyd ZE Murine models of obesity or reduced adiposity are a valuable resource for understanding the role of adipocyte dysfunction in metabolic disorders. Primary adipocytes grown in culture and derived from murine adipose tissue are essential 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 stem cells along with a protocol for inducing adipogenesis in this cell population. PMID: 21082392 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Isolation of Human Adipose-Derived Stem Cells from Lipoaspirates. Methods Mol Biol. 2011;702:17-27 Authors: Yu G, Floyd ZE, Wu X, Halvorsen YD, Gimble JM 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. PMID: 21082391 [PubMed - as supplied by publisher] | | | | | | | | | | | | | | | | | | | | | | | | | Isolation and Culture of Rhesus Adipose-Derived Stem Cells. Methods Mol Biol. 2011;702:3-16 Authors: Gagliardi C, Bunnell BA Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Rhesus monkeys are physiologically and phylogenetically similar to humans, and they and their cells are valuable for biomedical research and evaluation of preclinical therapies. Here, we describe methods for the isolation, culture, and differentiation of rhesus adipose-derived stem cells (rASCs). PMID: 21082390 [PubMed - as supplied by publisher] | | | | | | | | | |  | |  | |  |
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