Identification, Isolation and Expansion of Adult Stem Cells

London
Friday, 12 May 2006

Identification, Isolation and Expansion of Adult Stem Cells
Friday, 12 May 2006 09:15 - 16:30

Birkbeck College
Basement Lecture Theatre
43 Gordon Square
London
WC1H 0PD
United Kingdom

Map and Directions

For many years transplantation protocols have utilised haemopoietic tissues containing adult stem cells. Subsequent progress in scientific and clinical research characterized the cells responsible for haemopoietic reconstitution. Recently, normal and cancer stem cells have been identified in a variety of tissues. Our ability to manipulate these stem cells and/or their progenitors should lead to exciting developments in tissue engineering, tissue regeneration, therapeutic intervention as well as facilitating functional and molecular studies. Talks from leaders in the field will highlight identification and isolation of normal and cancer stem cells; current protocols for their growth and their potential application in cell based therapies�� �V Dr Erica De Wynter, University of Leeds �V the Meeting��s Chair

09:00 �V 09:30 Registration �V Tea, coffee and biscuits

09:30 �V 09:40 Introduction by the Chair: Dr Erika de Wynter, University of Leeds


09:40 - 10:10
Regulation of human breast stem cells - Dr Rob Clarke, Breast Biology Group, University of Manchester, Christie Hospital (NHS) Trust.
We have developed methods for the isolation and characterisation of human breast epithelial stem cells based on those previously used for haematopoietic stem cells.  We have also adapted neural stem cell culture methods for the culture and investigation of the regulation of normal and malignant breast cells. Our studies indicate the importance of the Notch receptor signalling pathway in breast stem cells and cancer.  We are also using shRNA libraries to discover novel breast stem cell regulatory pathways.

 
10:10 �V 10:40 Phenotypic and Functional Characterization of Mouse Mammary Epithelial Stem Cells - Dr John Stingl - Stemcells Technologies
The frequency and phenotypic properties of mouse mammary epithelial stem cells is poorly understood. To address these issues, we combined multi-parameter cell sorting and limiting dilution transplant analysis into cleared mammary fat pads to characterize a rare subset of adult mouse mammary cells that are able to regenerate the mammary epithelium. Our results indicate that there are approximately 1,400 mammary stem cells in each inguinal mammary gland of a young adult female mouse and that these cells have a CD45-/Ter119-/CD31-/CD24med/CD49fbright phenotype and are not quiescent. Mixing experiments using marked cells and single cell transplants demonstrate that a single cell can engraft a cleared fat pad. Transplantation of cells harvested from primary outgrowths into secondary fat pads demonstrate that these stem cells have the ability to undergo up to 10 symmetric self-renewal cell divisions, thereby fulfilling the requirements of a mammary stem cell.

10:40 �V 11:10 The Diversity of Neural Stem Cells - Professor Jack Price - MRC Centre for Neurodegeneration Research, Institute of Psychiatry, London

Neural Stem Cells can be isolated from Adult or Fetal brain, or derived from embryonic stem cells.  Though these different sources of cells share seminal stem cell properties, they differ in other aspects of their phenotype.  Importantly, some of these isolates have the capacity to repair the damaged nervous system, for others this is less certain.  How are we to view this diversity of neural stem cell populations?  In this talk, I will present our attempts to correlate cell and molecular phenotypes, and to understand the mechanisms that drive neural stem cell diversity.

 
11:10 �V 11:40 Morning coffee/tea break in the exhibition hall

11:40 �V 12:10 Galectin -1 �V A way to repair muscle fibres? - Professor Diana Watt �V Brighton and Sussex Medical School

The finding of a cell within skin that is capable of entering the myogenic lineage under certain conditions prompted us to identify a candidate factor for this conversion. Having identified the factor as galectin-1 we have gone on to determine the importance of galectin-1 in muscle determination and differentiation and how it could enhance muscle repair and regeneration following trauma and disease. Since these initial experiments we have begun to dissect out the role of galectin-1 in muscle and start to determine how it could be used to convert undifferentiated cells to the myogenic lineage thereby finding a therapeutic use for the molecule in enhancing myogenesis.

12:10 �V 12.40
Isolation, characterization and use of human AC133 progenitor cells for vascular tissue repair
-
Dr Pauliina Lehtolainen, University College London

Talk includes isolation, characterisation and knowledge of AC133 cells and their lineage commitment towards endothelial cells and in vivo experience of their usage in tissue repair.


12.40 �V 13:10 Application of a defined stem cell population to the treatment of patients with liver disease - Dr Myrtle Gordon, Department of Haematology, Imperial College Faculty of Medicine, London

We have identified a subpopulation of stem cells in adult human haemopoietic tissue. These cells have been characterised and their in vitro growth and differentiation have been documented. A phase I clinical trial has investigated their safety and efficacy in patients with liver disease. Early indications of clinical efficacy warrant further clinical studies.

 
13:10 - 14:10 Lunch in the exhibition hall

14:10 �V 14:40 Complexity of the Human Haematopoietic Stem Cell compartment - Daniel Pearce, Cancer Research, UK

Daniel Pearce will summarise the various methods available for haematopoietic stem cell identification in both murine and human tissues. He will then go on to explain the way that we assess haematopoietic stem cells in the murine and human setting. The concept of the leukaemic stem cell will be introduced. He will present some of the work that has been performed in the laboratory using the aforementioned techniques. Specifically, he will present phenotyping analysis of cells identified by pump or enzyme activity, analysis of the phenotype of the cell responsible for the propagation of leukaemia and other related projects.

14.40 �V 15.10 Identification and isolation of Human Mesenchymal Stem Cells - Dr. Elena Jones - Leeds University

15:10 �V 15.40 Nonhematopoietic Stem Cells and their Potential for Cell Therapy - Susan Donath, Miltenyi Biotec

Throughout the whole life cycle, the human body maintains a supply of adult stem cells that are able to proliferate and differentiate into mature cells of multiple hematopoietic and nonhematopoietic (NH) lineages. These stem cells have the potential to revolutionize tissue regeneration therapies. the use of defined stem cell populations from the outset, in addition to reproducibility of data and standardization of protocols are key to unraveling the full potential of stem cells. Therefore, the establishment of efficient and reproducible procedures for the isolation of defined populations of stem cells for the implementation of reliable and predictable therapies is an important first step.

 

This presentation will give an overview about recent findings in research on stem cell from hematopoietic and nonhematopoietic sources and their regenerative potential.

 

Furthermore, it will be discussed what attempts have been made to isolate homogeneous populations of marrow stromal cells (MSCs) - nonhematopoietic stem cells from bone marrow - using different positive selection markers such as CD133, CD271 (LNGFR), Stro-1, CD117, CD105, and anti-fibroblast antigen. Furthermore, standardization of the cultivation and quality control of cultivated MSCs will be discussed.

 

Additionally, an introduction into the MACS (Magnetic Activated Cell Sorting) Technology, a method to separate specific cell populations by immunomagnetic labeling, will be given.

15:40 �V 16:10 Afternoon Tea/Coffee in the exhibition hall

16.10 �V 16:40 Prostate cancer stem cells - Dr AT Collins, Univeristy of York

There is now increasing evidence in some malignancies that tumour cells are organised as a hierarchy originating from rare stem cells that are responsible for maintaining the tumour. 

We have isolated tumour stem cells from patients with prostate cancer using the cell surface markers integrin ��2��1 and CD133. The stem cell population can be expanded and maintained in culture and display stem cell-like properties, such as self-renewal (they are capable of generating new clones containing additional stem cells), and they have the potential to regenerate phenotypically mixed populations of non-clonogenic cells present in the original tumours. 

16:40 �V 17:10 A Population Of Putative Stem Cells From Human Prostatic Epithelial PrecursorsTransplanted Into Nod-Scid Mice - R. P.Revoltella Institute of Biomedical Technologies, CNR, Pisa, Italy

An enriched population of human epithelial progenitors ( p63 , high m. w. CK (clone 34Eβ12)) was isolated from benign prostatic biopsies, expanded on feeder NIH.3T3 fibroblasts and then transplanted into nod-scid mice. Positive control mice received human CB-CD133 cells, while negative control mice only saline. After 7d and 30d, by PCR ( for HLA.DQα1 and 11 specific  microsatellites (CODIS)) human DNA was detected in all engrafted mice in different organs, but never in negative control mice. By FISH, distribution was restricted to areas known as stem cell niches. These findings suggest that prostatic epithelial cells may contain putative stem cells.

 


17:10 - 17:15 Close

 

Contact Details

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    Sally Wheatland
    Euroscicon
    PO Box 49717
    London
    N20 8WH.

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