Maintaining Stem cells and the regulation of their differentiation

Welwyn Garden City , Hertfordshire
Friday, 27 April 2007

Maintaining Stem cells and the regulation of their differentiation
Friday, 27 April 2007 09:00 - 17:00

BioPark Hertfordshire
Broadwater Road
Welwyn Garden City
Hertfordshire
AL7 3AX
United Kingdom

Map and Directions

"Much of the promise of stem cell biology and tissue engineering depends on the ability to maintain and differentiate cells into tissues in a robust and reproducible manner. We are only just at the beginning of understanding how to control cell growth in vitro and produce cell types that are of value to basic research, pharmaceutical development and potential therapeutic applications. This meeting is designed to bring together experts in the field to share their experiences of developing innovative ways to create a more realistic environment for cell growth and differentiation. There are many physical factors that are often taken for granted during the culture of cells, including the topography and nature of the substrate, oxygen tension, temperature, etc. Furthermore, the addition of exogenous factors to the culture medium and the development of co-culture models are of importance to more closely mimic the conditions cells experience in vivo. The aim of this symposium is to therefore recognise that the control of cell differentiation requires an in-depth understanding of the growth conditions and signals cells require in order to differentiate in a particular manner. This will be exemplified by a carefully selected panel of speakers who will share their experiences of controlling cell growth and differentiation using a range of alternative approaches"
Chairman:  Dr Stefan Przyborski, School of Biological and Biomedical Science, University of Durham



09:00 – 09:45 Registration - Tea/Coffee

09:15 – 09:35 *Tour of the BioPark by William Sprigings (Marketing Director) or Steve Read (General Manager)

09:45 – 10:00 Introduction by the Chair:
Dr Stefan Przyborski, School of Biological and Biomedical Science, Durham University, UK

10:00 – 10:30 Development of Enabling Technology to Control Cell Growth and Function In Vitro
Dr Stefan Przyborski, School of Biological and Biomedical Science, Durham University, UK
Through a collaboration working with Dr Neil Cameron also within the Centre for Bioactive Chemistry, we have developed novel apparatus for three-dimensional (3D) cell growth in vitro. Biotechnologists routinely grow cells on flat two-dimensional (2D) tissue culture plastic. This is far removed from the complex 3D environment cells experience during the differentiation of tissues in the embryo. We have used emulsion templating to produce 3D scaffolds manufactured from polystyrene – the same material of normal culture plastic-ware. Furthermore, such materials have been modified to fit into existing cell culture welled plates and dishes. We have exemplified the use of this technology by growing a range of alternative cell types on these surfaces. Cells grow within the 3D materials and we have evidence that cell differentiation is significantly enhanced on 3D substrates compared to 2D surfaces.

10:30 – 11:00 Development of co-culture system to enhance haematopoietic differentiation by ES cells
Dr Lesley Forrester, University of Edinburgh, Scotland
We have shown that exposure of differentiating ES cells to the primary aorta-gonadal mesonephros (AGM) microenvironment results in a significant increase in the number of ES-derived haematopoietic progenitors. Co-culture of ES cells on a stromal cell line derived from the AGM region also increases haematopoietic activity and will be useful in the identification of haematopoietic inducing growth factors. We will also present results using a novel pharmacological induction system to assess the role of the SCF/KIT signalling pathway during ES cell differentiation.

11:00 – 11:20 Morning Tea/Coffee and Poster Viewing

11:20 – 11:50 Effects of the microenvironment on ES cell differentiation
Dr Anne Bishop, Imperial College, London, UK
The differentiation potential of ESCs is theoretically unlimited, with formation of the three germ layers (ectoderm, mesoderm, endoderm) as first step. The most common method to achieve this is the differentiation of ESCs into three-dimensional structures in suspension, the embryoid bodies (EBs), although some groups are also using monolayer culture to try to control differentiation more. Cells within EBs can be further differentiated into a variety of committed cell types, although the resultant cultures inevitably contain multiple lineages/phenotypes. Means to upregulate the differentiation of specific phenotypes include medium supplementation, co-culture with cells/tissues, cell extract-based re-programming and genetic modification.

11:50 – 12:10 The maintenance of high levels of glycolysis by mesenchymal stem cells cultured under normoxic conditions
Mr Girish Pattappa, Queen Mary University of London, UK
The proliferation and differentiation of mesenchymal stem cells is influenced by the oxygen tension. To understand this effect and predict cell behaviour within 3D culture environments, this study measured the oxygen consumption, glucose consumption and lactate production. The mean consumption rate per cell for human mesenchymal stem cells ranged from 44.6 - 93 fmoles/hr/cell at passage 3. There was no significant difference in the oxygen consumption rate between P2-P4. The production of lactate suggests that these cells are preferentially glycolytic, despite the presence of oxygen, a phenomenon known as the Warburg effect.

12:10 – 12:40

Human CB-CD.133 Cells Injected TO Nod-Scid Mice Made Deaf After Ototoxi Treatment Provide Conditions For The Resumption Of The Inner Ear Structure
R. P. Revoltella, Institute of Biomedical Technologies, C.N.R., Pisa, Italy
We investigated whether human CD133 stem cells (HSC) engraft the cochlea and contribute to the restoration, in vivo, of the organ of Corti in nod-scid mice after ototoxic treatment with kanamycin and noise. By PCR, we tested the presence of HLA.DQa1, used for the traceability of the engrafted cells, demonstrating that the HSC injected i.v. migrated to various tissues of the host, including the organ of Corti. In oto-injured mice, HSC contributed to the stimulation ex novo of the morphological recovery of the organ of Corti, while the control oto-injured mice that were not transplanted, remained seriously damaged. Partial functional recovery was demonstrated after transplantation. Dual color-FISH- analysis provided further evidence of the positive engraftment in the cochlea , including the organ of Corti, as well as in different mouse tissues, and also revealed a small proportion of heterokaryons probably derived from fusion of donor with endogenous cells.

These observations offer the first evidence that, HSC migrating to the cochlea and contributing to the organ of Corti’s structure restoration, may provide conditions for the resumption of the inner ear.

12:40 – 13:00 Tour of the BioPark

13:10 – 14:00 Lunch and Poster Viewing

14:00 – 14:30 BD MatrigelTM - An Ideal Surface for Maintaining Human Embryonic Stem Cells
Dr Peter Weiser, European Product Manager, , BD Biosciences
Embryonic stem (ES) cells have an infinite capacity for self-renewal and are pluripotent (i.e. they can form all the cell types that make up the human body). A great deal of hope is associated with the potential application of human ES cells in cell therapy and regenerative medicine. Initial human ES cell derivation and culturing techniques originated predominantly from methodology developed for mouse ES cells, and human ES  cells therefore were cultured on mouse embryonic feeder (MEF) cells. While mouse feeder cells have proven to be a robust surface for long-term culture of human ES cells, they also present a number of severe limitations especially if human ES cells are to be routinely used in the clinic. For example, there are increasing concerns about xenogeneic contamination of human ES cells grown on mouse feeder cells. For these and other reasons, efforts have been initiated to develop feeder-free conditions for culturing human ES cells. BD MatrigelTM is presented as a surface for MEF-free culture to maintain human ES cells in an undifferentiated state.

14:30 – 15:00 Effect of culture temperature on mesenchymal stem cell viability and potency
Dr A.M.Scutt, University of Sheffield
Mesenchymal stem cells (MSC) can adopt all mesenchymal phenotypes and because of this are considered attractive as candidates for Tissue Engineering or Cell Therapies. However, unlike other stem cell types, MSC are not immortal and do not express telomerase. Furthermore, for therapeutic purposes they will be isolated from aged individuals and will suffer oxidative stress during expansion in vitro leading to senescence and compromised engraftment potential. We have investigated the effect of, among other things, culture temperature on MSC oxidative stress and have found that a modest temperature reduction minimises oxidative damage and maintains proliferation and differentiation potential.

15:00 – 15:30  Astrocyte signals influence adult stem cell potential.
Alexis Joannides, University of Cambridge, UK

15:30 – 16:00 Afternoon Tea/Coffee and Last Poster Viewing

16:00 – 16:30 Inhibition of neuritogenesis by adult neuroprogenitors grown on MAG-coated substrates
Dr Carla Mellough, Durham University
In the injured adult brain, myelin-associated inhibitors are expressed before the glial scar has formed and bind to the p75NTR-NgR complex on regenerating neurons causing growth cone collapse and axonal retraction. We have assessed the effects of myelin-associated glycoprotein (MAG) on neurite outgrowth in a neuronal population derived from adult hippocampal progenitors. We show that MAG does not alter progenitor cell fate but significantly attenuates neurite outgrowth from differentiating neurons. We demonstrate that this effect can be partially overcome by activation of neurotrophin, cAMP and PKA pathways or Rho-kinase suppression. Furthermore, combining treatments elicits enhanced neurite outgrowth under myelin inhibition.

16:30 – 17:00 The Power of Guava Technologies
Dr Paul Wheeler, Guava Technologies, UK

17:00 - 17:15  Chairman’s summing up & close.

*Please note that a tour for exhibitors is scheduled for 2:15- 2:45

 Please email abstracts for poster presentation to abstracts@euroscicon.com.  All accepted abstracts will be published in the meeting proceedings

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