|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
|Stem Cell Derived Human Neurons in CNS Drug Discovery|
L Jackson, C Verastegui, C Wells, D Pau, A Nunn, A Finucane, M Lynch, S Boldt, L Gerrard, S.J. MacKenzie
Central nervous system disorders affect many people worldwide. To increase the quality and number of drugs available for treatments of CNS disorders such as Alzheimer’s, schizophrenia and anxiety, the success rate of CNS drug discovery programmes must be improved. The challenge in drug discovery is to develop more relevant assay systems to identify lead compounds for further development. Cell based screening assays currently used for CNS drug development involve primary cells or commercially
|Current status and prospects of the commercial storage of adult stem |
Alexander E. Rodin, Anatoly N. Gromyko, Alexander B. Smolyaninov
The authors present a result of survey among 300 residents of the Russian Far East on their potential willingness to store cord blood stem cells of their newborns and/or their own gametes for the future needs. The current status of the cord blood banking system in Russia is also described.
|Modular Glass Chip System for the Acquisition of the Electric Activity and Physiological Parameters of Differentiated Stem Cells|
P. J. Koester, J. Sakowski, S. M. Buehler, C. Tautorat, H. Altrichter, W. Baumann and J. Gimsa
The EU chemical policy REACH, effective on June 2007, regulates the registration, evaluation and authorization of chemicals. Investigations on the effects of neurotoxic and developmental-neurotoxic substances are required by European and US-American test guidelines. However, at the moment these tests are based on animal experiments contradicting society claims to abolish animal experiments and to implement the 3R-principle. The resulting problem is… How to pair REACH and the 3Rs?
|Changes in the Gene Expression of mRNA Transcripts for Insulin like Growth Factor, their receptor and Facilitative Glucose Transporter in IVM Oocytes|
S.C. Gupta, Neelam Gupta and Alok Pandey
The cDNA libraries from single oocytes and pre-implantation buffalo embryos from 2 cell stage to blastocyst were established. Relative expression studied with RT-PCR of IGF-I showed an increase at 12h and decline at 24h of IVM oocytes. IGF-IR was expressed at cleavage stages to blastocyts. Glut-I was expressed in IVM oocytes and SCNT embryos at all stages. Gene expression of IGF-I, IGF-IR and Glut-I plays an important role in SCNT embryo production.
|Development of a Lab-on-a-Chip for the Characterization of Human Cells |
Richter, L., Stepper, C., Mak, A., Brückl, H. and Ertl, P.
Cell chips are developed to continuously monitor mammalian cell population dynamics in a non-invasive manner. In the presented work we describe the design, fabrication and characterization of a lab-on-a-chip for quantitative cell analysis.
|CyTRAK™ Probes: Novel Nuclear and Cytoplasm Discriminators Compatible with GFP-Based HCS and HTS Assays|
RJ Errington, LH Patterson, R Edward and PJ Smith
Image-based high-content screening assays, demand solutions for image segmentation and cellular compartment encoding to track critical events – for example those presented by GFPreporters within cell cycle tracking and GPCR translocation assays. We have designed nuclear and cytoplasm discriminator CyTRAK™ probes - spectrally compatible with all variants of GFPreporters offering new solutions in cytometry.