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Clinical collaborations lead to patient therapies As one of the foremost scientists in the rheumatology field and the development of chondrogenic cultures from BMSCs, Professor Hollander works closely with clinical teams to develop patient-specific cartilage autografts. These are generated by extracting cartilage cells (rather than BMSCs) from the patient and culturing them to provide autologous chondrocytes, which are then seeded into a three- dimensional biodegradable material (derived from the total esterification of hyaluronan with benzyl alcohol and constructed into a non-woven configuration). These engineered grafts are then placed at the site of the cartilage injury, often without the need to glue or suture them in place. Furthermore, the procedure does not necessitate open surgery since a miniarthrotomy is usually sufficient. Once in place, the graft quickly integrates with the patient’s existing tissues, providing good collagen composition and integration with the underlying bone. The autograft technique provides several distinct advantages, namely less stressful surgical procedures and perhaps more importantly, the lack of any immune response. This is a major advance over allografts, which require the use of powerful immunesuppressing drugs for extended periods post-transplant. The first ever bio-engineered tracheal graft Last summer, Professor Hollander received a request for help from a friend and colleague, Professor Martin Birchall, a surgical professor at the University of Bristol. A patient of Dr Birchall’s had suffered serious damage to her trachea as a result of contracting tuberculosis (TB). The patient, was a young mother whose only chance of survival at the time was to have one lung removed, which would have seriously affected her quality of life and her ability to look after her children. After much discussion, Professor Hollander and his team very quickly set to work adapting their existing osteoarthritis based protocols to enable Professor Birchall to grow a large population of chondrocytes derived from the patient’s BMSCs. A section of human trachea was donated for use as a scaffold on which the new tissue could be grown. The trachea was stripped of the donor’s cells, leaving a trunk of non- immunogenic connective tissue onto which the chondrocytes were seeded. This seeding process used a novel bioreactor developed at the Politechnico di Milano, Italy, which provided the right environment for the cells to form the cartilaginous part of the trachea within four days of seeding. The graft was then lined with epithelial cells and transplanted into the patient, who responded very quickly to the new airway section, without any sign of rejection (no antibodies to the graft were found). Subsequent biopsies have shown that the new section is fully integrated with the existing airway and is fully supplied with blood vessels. She is now able to live life as if she had not been struck down with TB, a result that would never have been possible if her lung had been removed. Discussion Stem cell based-therapies have promised huge changes in the treatments of many diseases and disorders, but much research is still required to ensure safety and consistency before they can be applied more extensively. Prof Hollander and his colleagues at the Department of Cellular & Molecular Medicine at the University of Bristol have been investigating the fundamental principles governing the differentiation of bone marrow stem cells into chondrocytes – the source of cartilage. Through this research they aim to further improve the processes used to generate chondrocyte-based autografts, which have already started to prove their value in the treatment of cartilage damage. Throughout their pioneering research, Professor Hollander’s team has come to rely on the dependability and functionality of a broad array of standard and advanced laboratory equipment specifically designed to provide the highest quality and reliability in the cell biology laboratory. As a result of their dedicated work, Professor Hollander’s team was able to take part in the amazing feat of the first ever bio-engineered tracheal graft. Their work has enabled this patient to regain an amazing quality of life following a life-threatening condition while increasing the drive among researchers and clinicians to more expansive use of stem cell based therapies. Feature article Professor Anthony Hollander, ARC Professor of Rheumatology & Tissue Engineering in the Department of Cellular & Molecular Medicine at the University of Bristol, UK. (Image courtesy of Dr Sally Dickinson, University of Bristol). Figure 1. Adherent human adult bone marrow stem cells in culture. (Image courtesy of Dr Sally Dickinson, University of Bristol) Figure 2. Tissue engineered cartilage produced from bone marrow stem cells (Macroscopic Appearance). (Images courtesy of Dr Sally Dickinson, University of Bristol). 21
Article Choosing the Right Centrifuge for Your Application Ms. Goodman – Sample Preparation and Separations Applications Product Manager, <strong>Thermo</strong> <strong>Fisher</strong> Scientific Inc Figure 1–Superspeed centrifuges are an excellent choice for multiuser, multiprotocol environments, offering high capacity, high g-forces, and a broad range of rotors and accessories. With so many recent advances in both science and technology, it is wise to educate yourself about the wide range of centrifuge options now available. Following are some key questions that can be used to determine the type of centrifuge that will best meet your needs: 1. What applications and protocols will the centrifuge be used to support? 2. What are the maximum and minimum g-force (relative centrifugal force, RCF) and volume requirements? 3. How many tubes or samples must be processed in a run, shift, or day? 4. What types of sample formats will the centrifuge need to support (i.e., microplates, blood collection tubes, disposable conical tubes)? 5. What type of rotors will be needed to support your applications (i.e., fixed angle, swinging bucket)? 6. How many people will be using the centrifuge? 7. Is versatility important? That is, do you anticipate the need for a broad range of protocols and multiple users, or will you be performing the same standardized protocol day after day? 8. Do you have any space restrictions, such as benchtop or floor space only? 9. Do you have special needs such as process traceability, user lock-out, or biocontainment? 10. What is your budget? Once you have answered these questions, you should have a better picture of your centrifuge requirements. It is also helpful to review the basic types of centrifuges to make sure you know which category best fits your needs. Floor model or benchtop? Centrifuges are generally classified as either floor-standing or benchtop models. The style you choose is typically determined by performance requirements, available space, and budget.Floor-model centrifuges free up bench space and are often chosen for either high-speed or high-capacity sample processing. Within the floor-model category there are superspeed centrifuges, ultracentrifuges, and low-speed centrifuges.Benchtop centrifuges offer versatility and convenience, and can be equipped to accommodate a broad range of needs, making them a cost-effective solution for many laboratories. Benchtop platforms include general-purpose centrifuges, micro- centrifuges, small clinical centrifuges, cell washers, and high-speed models. Choosing the right floormodel centrifuge Superspeed centrifuges – If you are looking for high capacity, high g-force and versatility, a superspeed centrifuge (Figure 1) is probably the best choice. Many superspeed models offer a choice of up to 40 rotors, making them an excellent solution for core laboratories performing general preparative applications such as whole cell separations, protein precipitation, tissue culture, subcellular isolation (i.e., Golgi bodies, ribosomes), plasmid preps, and DNA/RNA separations. Superspeed centrifuges are also the best option for multiuser, multiprotocol environments. Their versatility enables researchers to step into new, cutting-edge technologies without purchasing a dedicated centrifuge for one specific application. Ultracentrifuges – If your application calls for g-forces of up to 1,000,000 × g, you will need an ultracentrifuge. These extremely powerful centrifuges support sample volumes up to 250mL. Within the ultracentrifuge line there are two platforms: full-size floor models, which support g-forces of up to 802,000 × g and volumes up to 250mL; and micro-ultracentrifuges, which support g-forces of above 1,000,000 × g and microvolume samples up to 13.5mL. Common ultracentrifuge applications include the separation of virus particles; DNA, protein, or RNA fractionation; as well as lipoprotein flotation. Density and size gradient 22
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