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Supernatant of estrogen-treated human umbilical cord blood derived hematopoietic stem cells under extremely hypoxia attenuate experimental: spinal cord injury
S24
Poster abstracts
73 SUPERNATANT OF ESTROGEN-TREATED HUMAN UMBILICAL CORD BLOOD DERIVED HEMATOPOIETIC STEM CELLS UNDER EXTREMELY HYPOXIA ATTENUATE EXPERIMENTAL: SPINAL CORD INJURY S Chen1, W Lo2, H Chang2, C Wu1 1 Chi Mei Medical Center, Tainan, Taiwan, 2Stem cell Research Center, Health Banks Co., Ltd., Taipei, Taiwan From our previous studies, both of human umbilical cord blood derived hematopoietic stem cells (hUCBHSCs) transplantation and estrogen (E2) administration can improve hindlimb motor dysfunction after experimental spinal cord injury (SCI) respectively. It indicated these two agents may have the potentials of anti-inflammation, vasculogenesis and neurogenesis. However, there is still persisted graft-versus-host disease (GVHD) in hUCBHSCs transplantation therapy. Therefore, we designed 17bestradiol (E2)-stimulated hUCBHSCs cultured in HSC medium under extreme hypoxic (0.1% O2) for three days. Three days later, the conditioned medium (CM) harvested from supernatant (cell free). Rats were divided into six groups: (1) sham operation (laminectomy only); (2) SCI + 0.5 cc saline, iv, N¼6; (3) SCI + 0.5 cc HSC medium, iv, N¼6; and (4) SCI + 0.5 cc CM1 (HSCs-1106+ HSC medium), iv, N¼6; (5) SCI + 0.5 cc CM2 (E2+HSCs-1106+ HSC medium), iv, N¼6; and (6) SCI + 0.5 cc CM3 (E2+ HSC medium), iv, N¼6. SCI was induced by compressing the spinal cord (T8-T9) for 1 min with an aneurysm clip calibrated to a closing pressure of 55 g. All the administered were injected immediately after SCI via the tail vein. Behavioral tests of motor function invented by Basso, Beattie, Bresnahan (BBB) scoring was detected at day 1 to 7 after SCI. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay was also conducted after SCI to evaluate spinal cord apoptosis. The Immunohistochemical staining of glial fibrillary acidic protein was conducted to evaluate astrogliosis. Enzyme-Linked Immunosorbent assay to analyze the amount of vascular endothelial growth factor (VEGF) and glial cell line-derived neurotrophic factor (GDNF) in the CM. It was found systemic administration of estrogen-treated HSC in HSCs medium under 0.1% O2 significantly attenuated the SCI induced hind limb dysfunction and spinal cord apoptosis. Both GDNF and VEGF could be detected significantly higher in the CM2. 74 WILL NOT BE PRESENTED 75 SUCCESSFUL USE OF PLERIXAFOR IN HARD TO MOBILIZE PATIENTS - FOLLOWING HIGH DOSE THERAPY ALL PATIENTS DEVELOPED FAST AND SUSTAINED ENGRAFTMENT WITH DURABLE CLINICAL RESPONSES D Josefsen, G Hetland, A Blystad, Y Fløisand, G Kvalheim Oslo University Hospital, Oslo, Norway Mobilization of autologous hematopoietic stem cells is most frequently performed using G-CSF alone. In our clinic we use G-CSF in combination with chemotherapy. In 15-20% of the patients insufficient numbers of CD34+ cells are harvested, and are regarded as poor-mobilizers. We have previously shown that mobilized patients appearing with low concentration of CD34+ cells (5-10 cells/mL) and recovered total white cell count >10109/L, can successfully harvest stem cells by addition of plerixafor. Moreover, poor mobilizing lymphoma patients have a less favorable prognosis than good mobilizers. Here, we have mobilized and harvested 33 poor-mobilizing cancer patients by addition of plerixafor. Furthermore, the 16 lymphoma patients were followed up after reinfusion of autologous stem cells with regard to engraftment as well as clinical response. The day prior to harvest of the 33 patients, the level of leukocytes was 21,8109 cells/L (median), and the CD34+ concentration was 5,5106/L (median). Following plerixafor injection, the concentration of CD34+ cells increased to 26,8106/L (median). The patients were then successfully harvested (median: 3,9106 CD34+cells/kg) with 1-2 days of apheresis. High dose therapy is a curative treatment for lymphomas. Therefore, we focused especially on the clinical outcome on these16 patients. Following high dose therapy, time to short term engraftment, defined by neutrophils >0.5109/L and thrombocytes >20109/L were 11 days (median) and 20 days (median). Moreover, in contrast to previous findings we have observed durable
responses with relapse free survival of 77% and overall survival of 93%, with a median observation time of 14 months. In conclusion, our findings show that addition of plerixafor in hard to mobilize patients allow them to proceed to high dose therapy. Moreover, the additional costs related to the use of plerixafor can be justified since lymphoma patients obtain similar event free and overall survival as those patients that were good mobilizers. 76 ESTABLISHING AN ALGORITHM TO ENSURE AN OPTIMAL YIELD OF MOBILISED PROGENITOR CELLS PG Dyson1, S Hiwase1, LB To1,2, I Lewis1,2 1 SA Pathology, Adelaide, Australia, 2Royal Adelaide Hospital, Adelaide, Australia Aim: Haemopoietic reconstitution post transplantation depends on dose of haemopoietic progenitor cells (HPC) infused. Collection of an adequate dose HPC depends on effective mobilisation of HPC from the marrow into the circulation. Poor mobilisation affects patient outcome and resource utilisation. To maximise HPC harvest we sought to optimise mobilisation and collection protocols by identifying developing an algorithm that would ensure collection of optimal numbers of HPC. Method: We reviewed data for 128 patients who underwent progenitor cell mobilisation and autologous transplantation in our institution in 2009-2011. For this study we defined the transplant CD34+ cell dose as being optimal (6106/kg for MM and 3106/kg for NHL), low (2 to 6106/kg in MM and 2 to 3106/kg in NHL) and poor (<2106/kg in MM and NHL). Results: The target CD34+ cell dose was achieved during the first mobilisation in 100/128 (78%) patients. Multiple mobilisation cycles were performed in 19/128 (15%) patients. CD34+ cell yield correlated with circulating pre CD34+ levels - Spearman r2 ¼0.51. An optimal dose could not be collected in patients who failed to reach CD34+/ml ¼ 15.
Collection details
Details Gender Male Female Age First mobilisation G-CSF Chemo + G-CSF First mobilisation dose Optimal Suboptimal Second mobilisation dose Optimal Poor Low No collection
Multiple Myeloma (MM)
Lymphoma (NHL)
45 35 60 (24 - 70)
31 17 55 (19 - 69)
12 68
2 46
66 14
34 14
2 5 3 -
4 2 2 1
Conclusion: In both patient groups a CD34+/ml < 15 predicted an optimal CD34+ cell dose would not be collected. For G-CSF mobilised patients if the day 5 CD34+/ml < 7 then collection of an optimal cell yield is unlikely. For chemotherapy ¼ G-CSF mobilized patients if the day 10 white cell count < 2 109/l then collection of an optimal cell dose is unlikely.
77 G-CSF PRIMED DONOR HAEMATOPOIETIC STEM CELL COLLECTIONS ARE ASSOCIATED WITH REDUCED VIABLE T CELL YIELD FOR DONOR LYMPHOCYTE INFUSION V Antonenas1, F Garvin1, K Yehson1, G Hansra1, D McCulloch2, E Blyth2, M Hertzberg2, D Gottlieb1,2 1 Sydney Cellular Therapies Laboratory, Westmead Hospital, Australia, 2Blood and Marrow Transplant Service, Westmead Hospital, Australia
Poster abstracts
73 SUPERNATANT OF ESTROGEN-TREATED HUMAN UMBILICAL CORD BLOOD DERIVED HEMATOPOIETIC STEM CELLS UNDER EXTREMELY HYPOXIA ATTENUATE EXPERIMENTAL: SPINAL CORD INJURY S Chen1, W Lo2, H Chang2, C Wu1 1 Chi Mei Medical Center, Tainan, Taiwan, 2Stem cell Research Center, Health Banks Co., Ltd., Taipei, Taiwan From our previous studies, both of human umbilical cord blood derived hematopoietic stem cells (hUCBHSCs) transplantation and estrogen (E2) administration can improve hindlimb motor dysfunction after experimental spinal cord injury (SCI) respectively. It indicated these two agents may have the potentials of anti-inflammation, vasculogenesis and neurogenesis. However, there is still persisted graft-versus-host disease (GVHD) in hUCBHSCs transplantation therapy. Therefore, we designed 17bestradiol (E2)-stimulated hUCBHSCs cultured in HSC medium under extreme hypoxic (0.1% O2) for three days. Three days later, the conditioned medium (CM) harvested from supernatant (cell free). Rats were divided into six groups: (1) sham operation (laminectomy only); (2) SCI + 0.5 cc saline, iv, N¼6; (3) SCI + 0.5 cc HSC medium, iv, N¼6; and (4) SCI + 0.5 cc CM1 (HSCs-1106+ HSC medium), iv, N¼6; (5) SCI + 0.5 cc CM2 (E2+HSCs-1106+ HSC medium), iv, N¼6; and (6) SCI + 0.5 cc CM3 (E2+ HSC medium), iv, N¼6. SCI was induced by compressing the spinal cord (T8-T9) for 1 min with an aneurysm clip calibrated to a closing pressure of 55 g. All the administered were injected immediately after SCI via the tail vein. Behavioral tests of motor function invented by Basso, Beattie, Bresnahan (BBB) scoring was detected at day 1 to 7 after SCI. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay was also conducted after SCI to evaluate spinal cord apoptosis. The Immunohistochemical staining of glial fibrillary acidic protein was conducted to evaluate astrogliosis. Enzyme-Linked Immunosorbent assay to analyze the amount of vascular endothelial growth factor (VEGF) and glial cell line-derived neurotrophic factor (GDNF) in the CM. It was found systemic administration of estrogen-treated HSC in HSCs medium under 0.1% O2 significantly attenuated the SCI induced hind limb dysfunction and spinal cord apoptosis. Both GDNF and VEGF could be detected significantly higher in the CM2. 74 WILL NOT BE PRESENTED 75 SUCCESSFUL USE OF PLERIXAFOR IN HARD TO MOBILIZE PATIENTS - FOLLOWING HIGH DOSE THERAPY ALL PATIENTS DEVELOPED FAST AND SUSTAINED ENGRAFTMENT WITH DURABLE CLINICAL RESPONSES D Josefsen, G Hetland, A Blystad, Y Fløisand, G Kvalheim Oslo University Hospital, Oslo, Norway Mobilization of autologous hematopoietic stem cells is most frequently performed using G-CSF alone. In our clinic we use G-CSF in combination with chemotherapy. In 15-20% of the patients insufficient numbers of CD34+ cells are harvested, and are regarded as poor-mobilizers. We have previously shown that mobilized patients appearing with low concentration of CD34+ cells (5-10 cells/mL) and recovered total white cell count >10109/L, can successfully harvest stem cells by addition of plerixafor. Moreover, poor mobilizing lymphoma patients have a less favorable prognosis than good mobilizers. Here, we have mobilized and harvested 33 poor-mobilizing cancer patients by addition of plerixafor. Furthermore, the 16 lymphoma patients were followed up after reinfusion of autologous stem cells with regard to engraftment as well as clinical response. The day prior to harvest of the 33 patients, the level of leukocytes was 21,8109 cells/L (median), and the CD34+ concentration was 5,5106/L (median). Following plerixafor injection, the concentration of CD34+ cells increased to 26,8106/L (median). The patients were then successfully harvested (median: 3,9106 CD34+cells/kg) with 1-2 days of apheresis. High dose therapy is a curative treatment for lymphomas. Therefore, we focused especially on the clinical outcome on these16 patients. Following high dose therapy, time to short term engraftment, defined by neutrophils >0.5109/L and thrombocytes >20109/L were 11 days (median) and 20 days (median). Moreover, in contrast to previous findings we have observed durable
responses with relapse free survival of 77% and overall survival of 93%, with a median observation time of 14 months. In conclusion, our findings show that addition of plerixafor in hard to mobilize patients allow them to proceed to high dose therapy. Moreover, the additional costs related to the use of plerixafor can be justified since lymphoma patients obtain similar event free and overall survival as those patients that were good mobilizers. 76 ESTABLISHING AN ALGORITHM TO ENSURE AN OPTIMAL YIELD OF MOBILISED PROGENITOR CELLS PG Dyson1, S Hiwase1, LB To1,2, I Lewis1,2 1 SA Pathology, Adelaide, Australia, 2Royal Adelaide Hospital, Adelaide, Australia Aim: Haemopoietic reconstitution post transplantation depends on dose of haemopoietic progenitor cells (HPC) infused. Collection of an adequate dose HPC depends on effective mobilisation of HPC from the marrow into the circulation. Poor mobilisation affects patient outcome and resource utilisation. To maximise HPC harvest we sought to optimise mobilisation and collection protocols by identifying developing an algorithm that would ensure collection of optimal numbers of HPC. Method: We reviewed data for 128 patients who underwent progenitor cell mobilisation and autologous transplantation in our institution in 2009-2011. For this study we defined the transplant CD34+ cell dose as being optimal (6106/kg for MM and 3106/kg for NHL), low (2 to 6106/kg in MM and 2 to 3106/kg in NHL) and poor (<2106/kg in MM and NHL). Results: The target CD34+ cell dose was achieved during the first mobilisation in 100/128 (78%) patients. Multiple mobilisation cycles were performed in 19/128 (15%) patients. CD34+ cell yield correlated with circulating pre CD34+ levels - Spearman r2 ¼0.51. An optimal dose could not be collected in patients who failed to reach CD34+/ml ¼ 15.
Collection details
Details Gender Male Female Age First mobilisation G-CSF Chemo + G-CSF First mobilisation dose Optimal Suboptimal Second mobilisation dose Optimal Poor Low No collection
Multiple Myeloma (MM)
Lymphoma (NHL)
45 35 60 (24 - 70)
31 17 55 (19 - 69)
12 68
2 46
66 14
34 14
2 5 3 -
4 2 2 1
Conclusion: In both patient groups a CD34+/ml < 15 predicted an optimal CD34+ cell dose would not be collected. For G-CSF mobilised patients if the day 5 CD34+/ml < 7 then collection of an optimal cell yield is unlikely. For chemotherapy ¼ G-CSF mobilized patients if the day 10 white cell count < 2 109/l then collection of an optimal cell dose is unlikely.
77 G-CSF PRIMED DONOR HAEMATOPOIETIC STEM CELL COLLECTIONS ARE ASSOCIATED WITH REDUCED VIABLE T CELL YIELD FOR DONOR LYMPHOCYTE INFUSION V Antonenas1, F Garvin1, K Yehson1, G Hansra1, D McCulloch2, E Blyth2, M Hertzberg2, D Gottlieb1,2 1 Sydney Cellular Therapies Laboratory, Westmead Hospital, Australia, 2Blood and Marrow Transplant Service, Westmead Hospital, Australia