|Intra-operative Bone Marrow Processing for Bone Regeneration|
E. Nedelcu MD, A. Jamali MD, L. Hill RN, J. Nolta PhD, D. Vierria RN, M. Hoze RN, D. Taylor MD PhD, C. Gresens MD, D. Dwyre MD
Concentrated stem cell products obtained from autologous bone marrow aspirate were previously shown to regenerate avascular necrosis of the bone and non-union fractures. The aspirate processing was performed using apheresis technology, however, a detailed protocol has not been published yet. The aim of our study was to develop a clinical protocol using the CobeSpectra apheresis device to process the bone marrow aspirate.
|The Expression and Activity of MMPs during Zebrafish (Danio rerio) Development|
N.D. Harris, D.H. Li, J.Y. Keow, C. Po, A. Hong, K. Herrmann, B.D. Crawford
Recent advances have made the zebrafish an attractive system for studying MMP activity. However, there are few commercially available antibodies against MMPs that are suitable for studies using zebrafish. The objectives of the study presented in the poster were to develop effective immunological reagents for the study of MMPs in zebrafish and to test commercially available narrow-spectrum fluorogenic MMPs substrates in in vivo zymography.
|Genetic profile of human embryonic stem cells (hESC) grown under different O2 concentrations|
Sonia Prado, David Montaner, Dario Melguizo,Rubén Moreno, Joaquín Dopazo and Miodrag Stojkovic.
The development of mammalian embryos during the early stages takes place in a hypoxic environment. After 11 weeks of pregnancy the mother’s blood begins to flow freely in the placental space increasing the partial pressure of oxygen [1-3]. Therefore, we hypothesized that in vitro growth of hESC in normoxic conditions (21% O2) may change the profile and differentiation potential of hESC.
|Induced Pluripotency with Endogenous and Inducible Genes|
The recent discovery that two partly overlapping sets of four genes induce nuclear reprogramming of mouse and human has opened up new possibilities for cell replacement therapies. Although the combination of genes that induce pluripotency to some extenet, Oct4 and Sox2 appear to be a prerequisite. The introduction of four genes, several of which have been linked with cancer, using retroviral approaches is however unlikely to be suitable for future clinical applications.
|Human Fetal Bone Cells Seeded in Hyaluronic Acid Gel As A Delivery System For Bone Engineering|
D.Tenorio, L.A.Applegate, S. Jaccoud, C.Scaletta, D.P. Pioletti
Human fetal bone cells seeded in a hyaluronic acid gel as a delivery system, can survive and proliferate within this gel, are positive for alkaline phosphatase and von Kossa staining, demonstrating the potential of these cells bone engineering.
|Development of a Bone Marrow Targeting Cell Therapy|
Thomas Kean*, Lori Duesler*, Tambet Teesalu**, Erkki Ruoslahti**, James E Dennis*
One major obstacle in stem cell therapy is poor cell engraftment. To address this issue, concurrent investigations were made to develop novel peptides targeting bone marrow and a method to transiently ‘paint’ targeting molecules onto cells. Painting was studied using a model palmitated peptide. Novel bone marrow targeting peptides were sought using in vivo phage display in a locally irradiated mouse model that has an internal control (untreated leg).
|Adaptive control of glycerol & methanol feeding in recombinant Pichia pastoris cultures: Impact on antibody titre|
Filipe Ataíde, Ana Raquel Ferreira, João J. Clemente, António E. Cunha, Rui Oliveira
Pichia pastoris processes for heterolougous protein expression are typically run in three phases: a batch phase, an exponential feeding fed-batch phase and a long oxygen transfer limitation (OTL) phase. The final protein titre is often limited by the oxygen availability in the OTL phase. Here a direct adaptive controller is designed for the OTL phase. The controller was validated in pilot P. pastoris cultivations expressing a single chain antibody fragment (ScFv). This work shows that the propos
|Critical factors for successful RNAi experiments in primary cells and hard to transfect cell lines|
Markus Zumbansen1, Nicole Spottke1, Sheila Offizier1, Allison St. Amand2, Devin Leake2, Ludger Altrogge1, Meike Weigel1, Sandra Domzalski1, Dietmar Lenz, and Herbert Müller-Hartmann1
The amaxa Nucleofector® Technology is a well established method for effective, non-viral transfection of any nucleic acid substrate into hard-to-transfect cells, especially suspension and primary cells. With the
Nucleofector® 96-well Shuttle® system high throughput applications such as siRNA-library screenings have become amenable for the first time in these cell types. This renders target validation and identification possible
in cell types highly relevant for biomedical research. Here we dis
|High Content Image Analysis using a Laser Scanning Microplate Cytometer|
Paul Wylie, Ben Schenker, Sarah Payne and Andrew Goulter
Microscope-based high-content instruments offer high optical resolution, however, the limited field of view afforded by their objective lenses can mean lengthy read times for some assays, especially where multiple image capture per well is required. Wherever possible, only a small percentage of the total number of cells present in a test well are analysed to keep plate read times at a minimum, which may not always be ideal. Microplate laser- scanning cytometers, such as TTP LabTech’s Acumen® eX3