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Aldehyde dehydrogenase, but not viability of CD34 cells, predicts potency and engraftment after cord blood transplantation
S14
Oral Abstracts
Hematopoietic stem cells (HSC) have the capability for multi-lineage differentiation and self-renewal. Bone marrow and blood HSC transplants have been utilized to replace diseased bone marrow for patients with cancers and blood disorders due to its ability to differentiate into all kinds of immune cells. Following HSC transplantation, patients are cytopenic until donor cell engraft and may have long-term immune defective with accrued mortality risk arising from engraftment failure, opportunistic infection and disease relapse. Thus, novel strategies to efficient expansion of HSC can accelerate post-transplant engraftment of HSC and immune reconstitution, leading to improved survival of patients undergoing transplantation. We have recently bioengineered a novel cytokine that is derived from the fusion of GM-CSF and IL-4 (named as GIFT4) and have made the remarkable observation that GIFT4 fusokine induces endogenous B cell expansion and triggers entirely novel B cell function on HSC. Delivery of GIFT4 protein robustly promotes robust HSC expansion in the bone marrow that is dependent on the presence of B cells. Administration of GIFT4 protein together with B cells rescued lethally irradiated mice from bone marrow failure and death. Adoptive transfer of GIFT4-treated B cells prevented mice from irradiation-induced death. We propose that GIFT4 as well as GIFT4-licensed B cells could serve as means to augment HSC engraftment in the setting of bone marrow and HSC transplantation for patients with hematological malignancy and blood disorders. 25 ALDEHYDE DEHYDROGENASE, BUT NOT VIABILITY OF CD34 CELLS, PREDICTS POTENCY AND ENGRAFTMENT AFTER CORD BLOOD TRANSPLANTATION K Shoulars, JD Troy, T Gentry, P Noldner, K Page, A Balber, J Kurtzberg Duke University Medical Center, Durham, North Carolina, United States Delayed engraftment and graft failure are barriers to the success of unrelated donor cord blood transplantation (UCBT). This is due, in part, to decreased potency of the infused cord blood unit (CBU). We previously demonstrated that post-thaw colony forming units (CFU) is a strong predictor of survival and engraftment after
UCBT. However, the usefulness of CFU is limited by time and variability of assay results. Recently Barker and colleagues showed that viability of the CD34 population of a thawed CBU product predicted the engrafting unit in double UCBT. However, an assay assessing potency of the CBU prior to final selection for transplantation would be optimal. We developed a potency assay performed on segment attached to a CBU, enumerating the content of ALDHbr [AldecountÒ], CD34+, CD45+, glycophorin A+ and viability (7-AAD+), with CFUs, on CBUs requested for human leukocyte antigen confirmatory typing. From March 2010August 2013, segments from 2766 CBUs were analyzed. The percentage of viable CD45+ cells expressing ALDHbr (r¼0.82) or ALDHbr/CD34+ (r¼0.72) correlated well with CFUs (n¼2730, both p<0.0001). In contrast, CFU correlated less well with the percent of viable CD45+ cells expressing CD34+ (n¼2727, r¼0.28, p<0.0001) or the overall viability of CD34+ cells (n¼2727, r¼0.13, p<0.0001). In a cohort of patients with malignancies receiving a single CBU transplant after myeloablative conditioning, patients receiving cords low in ALDHbr or CFU experienced delayed engraftment compared to patients receiving patients receiving cord with higher levels of either CFUs or ALDHbr cells. In contrast, the viable CD34 content of the transplanted CBU was not a significant predictor of engraftment delay. Therefore, ALDHbr, which highly correlates with CFU, may be superior to viability of the CD34 population as a post-thaw potency measurement for CBU. Assaying the segment allows for identification of CBUs with higher potency before final selection of a CBU for UCBT. 26 DELIVERING A CELL THERAPY G Kirby1,2, L Vandenpoel3,2, J Pinxteren3,2, R Short1,2, A Michelmore1,2 1 Mawson Institute, University of South Australia, Mawson Lakes, South Australia, Australia, 2Cooperative Research Centre for Cell Therapy Manufacturing, Mawson Lakes, South Australia, Australia, 3ReGenesys, Heverlee, Belgium Background: Chronic wounds are a growing clinical burden on healthcare systems and with increasing incidences of obesity, diabetes and an aging population this problem is set to snowball. Chronic wounds present a hostile environment and unique challenges. Cell therapies may offer a solution with a range of cell based approaches beginning to cross the rift from benchto-bedside. Supporting data suggests that the appropriate administration of stem cells can accelerate wound healing. The effectiveness of a therapy will depend not only on what we deliver but how we deliver it. Aims or Objectives: Our aim was to generate a method of delivering cells to wounds where injection may not be ideal. We achieved this by manufacturing a cell-laden bandage. This bandage was capable of supporting cell attachment without affecting phenotype. When placed onto a wound, the cells then leave this surface in preference for the wound. Materials and Methods: Plasma polymerisation was used to generate a range of functional surfaces on candidate dressings. Coating conditions were varied and these surfaces were then tested with multipotent adult progenitor cells (MAPC). An in vitro transfer assay was used to determine the migration of viable cells from the patch onto a wound model. Results & Discussion: By varying surface treatments we were able to optimise a range of surfaces that supported the attachment of MAPC and subsequently allow these cells to leave in preference for a model wound bed. Conclusions: There is mounting evidence showing that cell therapies are effective in the treatment of chronic wounds. In order to promote clinical uptake of these new approaches, we need to make the use and delivery of cell therapies simple and effective. Our approach aims to create a simple bandage that is non-traumatic and easily applied at the bedside.
27 COST OF STEM CELL-BASED TISSUE-ENGINEERED AIRWAY TRANSPLANTS IN THE UK: CASE SERIES E Culme-Seymour1, K Mason2, L Vallejo-Torres3, C Carvalho4, L Partington4, C Crowley5, N Hamilton5, E Toll6, C Butler6, MJ Elliott6, MA Birchall5, C Mason7, M Lowdell4 1 London Regenerative Medicine Network, London, United Kingdom, 24 Newark Street, E1 2AT, Centre for Cutaneous Research, Blizard Institute of Cell and Molecular Science, London, United Kingdom, 3Clinical Trials Unit, Gower Street, WC1E 6BT, University College London, London, United Kingdom, 4Paul O’Gorman Laboratory of Cellular Therapeutics, University College London, Department of Haematology, Pond St, NW3 2QG, University College London, Royal Free Hospital, London, United
Oral Abstracts
Hematopoietic stem cells (HSC) have the capability for multi-lineage differentiation and self-renewal. Bone marrow and blood HSC transplants have been utilized to replace diseased bone marrow for patients with cancers and blood disorders due to its ability to differentiate into all kinds of immune cells. Following HSC transplantation, patients are cytopenic until donor cell engraft and may have long-term immune defective with accrued mortality risk arising from engraftment failure, opportunistic infection and disease relapse. Thus, novel strategies to efficient expansion of HSC can accelerate post-transplant engraftment of HSC and immune reconstitution, leading to improved survival of patients undergoing transplantation. We have recently bioengineered a novel cytokine that is derived from the fusion of GM-CSF and IL-4 (named as GIFT4) and have made the remarkable observation that GIFT4 fusokine induces endogenous B cell expansion and triggers entirely novel B cell function on HSC. Delivery of GIFT4 protein robustly promotes robust HSC expansion in the bone marrow that is dependent on the presence of B cells. Administration of GIFT4 protein together with B cells rescued lethally irradiated mice from bone marrow failure and death. Adoptive transfer of GIFT4-treated B cells prevented mice from irradiation-induced death. We propose that GIFT4 as well as GIFT4-licensed B cells could serve as means to augment HSC engraftment in the setting of bone marrow and HSC transplantation for patients with hematological malignancy and blood disorders. 25 ALDEHYDE DEHYDROGENASE, BUT NOT VIABILITY OF CD34 CELLS, PREDICTS POTENCY AND ENGRAFTMENT AFTER CORD BLOOD TRANSPLANTATION K Shoulars, JD Troy, T Gentry, P Noldner, K Page, A Balber, J Kurtzberg Duke University Medical Center, Durham, North Carolina, United States Delayed engraftment and graft failure are barriers to the success of unrelated donor cord blood transplantation (UCBT). This is due, in part, to decreased potency of the infused cord blood unit (CBU). We previously demonstrated that post-thaw colony forming units (CFU) is a strong predictor of survival and engraftment after
UCBT. However, the usefulness of CFU is limited by time and variability of assay results. Recently Barker and colleagues showed that viability of the CD34 population of a thawed CBU product predicted the engrafting unit in double UCBT. However, an assay assessing potency of the CBU prior to final selection for transplantation would be optimal. We developed a potency assay performed on segment attached to a CBU, enumerating the content of ALDHbr [AldecountÒ], CD34+, CD45+, glycophorin A+ and viability (7-AAD+), with CFUs, on CBUs requested for human leukocyte antigen confirmatory typing. From March 2010August 2013, segments from 2766 CBUs were analyzed. The percentage of viable CD45+ cells expressing ALDHbr (r¼0.82) or ALDHbr/CD34+ (r¼0.72) correlated well with CFUs (n¼2730, both p<0.0001). In contrast, CFU correlated less well with the percent of viable CD45+ cells expressing CD34+ (n¼2727, r¼0.28, p<0.0001) or the overall viability of CD34+ cells (n¼2727, r¼0.13, p<0.0001). In a cohort of patients with malignancies receiving a single CBU transplant after myeloablative conditioning, patients receiving cords low in ALDHbr or CFU experienced delayed engraftment compared to patients receiving patients receiving cord with higher levels of either CFUs or ALDHbr cells. In contrast, the viable CD34 content of the transplanted CBU was not a significant predictor of engraftment delay. Therefore, ALDHbr, which highly correlates with CFU, may be superior to viability of the CD34 population as a post-thaw potency measurement for CBU. Assaying the segment allows for identification of CBUs with higher potency before final selection of a CBU for UCBT. 26 DELIVERING A CELL THERAPY G Kirby1,2, L Vandenpoel3,2, J Pinxteren3,2, R Short1,2, A Michelmore1,2 1 Mawson Institute, University of South Australia, Mawson Lakes, South Australia, Australia, 2Cooperative Research Centre for Cell Therapy Manufacturing, Mawson Lakes, South Australia, Australia, 3ReGenesys, Heverlee, Belgium Background: Chronic wounds are a growing clinical burden on healthcare systems and with increasing incidences of obesity, diabetes and an aging population this problem is set to snowball. Chronic wounds present a hostile environment and unique challenges. Cell therapies may offer a solution with a range of cell based approaches beginning to cross the rift from benchto-bedside. Supporting data suggests that the appropriate administration of stem cells can accelerate wound healing. The effectiveness of a therapy will depend not only on what we deliver but how we deliver it. Aims or Objectives: Our aim was to generate a method of delivering cells to wounds where injection may not be ideal. We achieved this by manufacturing a cell-laden bandage. This bandage was capable of supporting cell attachment without affecting phenotype. When placed onto a wound, the cells then leave this surface in preference for the wound. Materials and Methods: Plasma polymerisation was used to generate a range of functional surfaces on candidate dressings. Coating conditions were varied and these surfaces were then tested with multipotent adult progenitor cells (MAPC). An in vitro transfer assay was used to determine the migration of viable cells from the patch onto a wound model. Results & Discussion: By varying surface treatments we were able to optimise a range of surfaces that supported the attachment of MAPC and subsequently allow these cells to leave in preference for a model wound bed. Conclusions: There is mounting evidence showing that cell therapies are effective in the treatment of chronic wounds. In order to promote clinical uptake of these new approaches, we need to make the use and delivery of cell therapies simple and effective. Our approach aims to create a simple bandage that is non-traumatic and easily applied at the bedside.
27 COST OF STEM CELL-BASED TISSUE-ENGINEERED AIRWAY TRANSPLANTS IN THE UK: CASE SERIES E Culme-Seymour1, K Mason2, L Vallejo-Torres3, C Carvalho4, L Partington4, C Crowley5, N Hamilton5, E Toll6, C Butler6, MJ Elliott6, MA Birchall5, C Mason7, M Lowdell4 1 London Regenerative Medicine Network, London, United Kingdom, 24 Newark Street, E1 2AT, Centre for Cutaneous Research, Blizard Institute of Cell and Molecular Science, London, United Kingdom, 3Clinical Trials Unit, Gower Street, WC1E 6BT, University College London, London, United Kingdom, 4Paul O’Gorman Laboratory of Cellular Therapeutics, University College London, Department of Haematology, Pond St, NW3 2QG, University College London, Royal Free Hospital, London, United