DEVELOPMENT OF A NON-INVASIVE STRATEGY ENHANCING HAEMATOPOIETIC STEM CELL (HSC) POTENTIAL OF HUMAN UMBILICAL CORD CELLS
Lead Research Organisation:
University of Edinburgh
Department Name: Edinburgh Research Office
Abstract
Transplantations of bone marrow containing blood stem cells termed haematopoietic stem cells (HSC) have been used for several decades for efficient treatment of various blood diseases. However, due to shortage of appropriate donors, the use of this treatment is limited. Search for methods of efficient production of HSCs or alternative sources enriched for HSCs is one of the key goals of current haematology.
Umbilical cord (UC) blood is an easily accessible and potentially very important source of HSCs but the number of HSCs contained there is low, which limits its use for treatment of adults. The aim of this project is to develop a methodology based on previous development in our laboratory which would allow significant enhancement of HSC activity of the umbilical cord. HSCs are efficiently generated during embryonic development and we have previously identified a powerful source of HSCs in the developing embryo, now termed the AGM region. We have developed a novel technique, which allowed us to generate large numbers of HSCs from the AGM region in culture. Here we propose to use the AGM cells for enhancement of the HSC activity of the umbilical cord. We will co-culture cells from the AGM region with cell populations of human umbilical cord cells.
To standardise this approach and also to make it compatible with clinical settings we will derive immortalised cell lines from the human AGM region and test their inductive capacity on umbilical cord cells. We will assess the production of HSCs from the umbilical cord by the gold standard test using transplantations in which long-term engraftment of human blood cells will be assessed in recipient animals.
Umbilical cord (UC) blood is an easily accessible and potentially very important source of HSCs but the number of HSCs contained there is low, which limits its use for treatment of adults. The aim of this project is to develop a methodology based on previous development in our laboratory which would allow significant enhancement of HSC activity of the umbilical cord. HSCs are efficiently generated during embryonic development and we have previously identified a powerful source of HSCs in the developing embryo, now termed the AGM region. We have developed a novel technique, which allowed us to generate large numbers of HSCs from the AGM region in culture. Here we propose to use the AGM cells for enhancement of the HSC activity of the umbilical cord. We will co-culture cells from the AGM region with cell populations of human umbilical cord cells.
To standardise this approach and also to make it compatible with clinical settings we will derive immortalised cell lines from the human AGM region and test their inductive capacity on umbilical cord cells. We will assess the production of HSCs from the umbilical cord by the gold standard test using transplantations in which long-term engraftment of human blood cells will be assessed in recipient animals.
Technical Summary
Definitive haematopoietic stem cells (HSCs) are responsible for the daily generation of billions of mature lymphoid and myeloid cells. Transplantation of one HSC into a myeloablated animal results in a long-term multi-lineage haematopoietic reconstitution. This strong regenerative potential of HSCs is broadly exploited in bone marrow transplantations procedures in clinical settings. HSCs are generated during embryonic development. The aorta-gonad-mesonephros (AGM) region plays an important role in the initiation and expansion of HSCs as has been shown in vitro. We have recently developed a novel ?open? AGM in vitro system which enables the effective initiation and dramatic (150-fold) expansion of HSCs, and importantly allows integration of exogenous cells, in a 3-dimensional co-aggregate culture.
Human umbilical cord is an easily accessible source of HSCs, but it contains low numbers of HSCs which limit its value for clinical applications. While useful for children, transplantation into adult patients is significantly less efficient. It is likely that at birth the umbilical cord still represents a ?plastic? stage in haematopoietic development. Published evidence suggests that the umbilical cord endothelium is still haematogenic.
This proposal aims to develop non-invasive methods for enhancing the HSC potential of human umbilical cord cells compatible with clinical settings. We will use primary AGM cells and derive a library of immortalised stromal cell lines from the human AGM region. AGM primary stroma and cell lines will be co-cultured with selected cell populations from the umbilical cord, which are phenotypically equivalent to cells developing into HSCs in the AGM region using our novel co-aggregation protocol. Formation of human HSC from umbilical cord cells will be monitored by long-term repopulating assay using severely immunocompromised NSG mice.
Human umbilical cord is an easily accessible source of HSCs, but it contains low numbers of HSCs which limit its value for clinical applications. While useful for children, transplantation into adult patients is significantly less efficient. It is likely that at birth the umbilical cord still represents a ?plastic? stage in haematopoietic development. Published evidence suggests that the umbilical cord endothelium is still haematogenic.
This proposal aims to develop non-invasive methods for enhancing the HSC potential of human umbilical cord cells compatible with clinical settings. We will use primary AGM cells and derive a library of immortalised stromal cell lines from the human AGM region. AGM primary stroma and cell lines will be co-cultured with selected cell populations from the umbilical cord, which are phenotypically equivalent to cells developing into HSCs in the AGM region using our novel co-aggregation protocol. Formation of human HSC from umbilical cord cells will be monitored by long-term repopulating assay using severely immunocompromised NSG mice.