Post-thaw Viability of Cryopreserved Hematopoietic Progenitor Cell Grafts: Does It Matter?

Cell viability in peripheral blood progenitor cell (PBPC) grafts and its influence on the clinical course following transplantation was evaluated in 81 consecutive transplantations (72 autologous, 9 allogeneic) performed in patients with hematological diseases. Viability of cells in PBPC grafts immediately upon collection was 98.6±3.5%, after addition of dimethyl sulfoxide (DMSO) 73.3±21.8%, and post-thaw 65.2±16.1%. It did not differ significantly between patients with different diagnoses, gender, age, type of priming used, dose of G-CSF administered or number of CD34+ cells collected. However, grafts stored for more than 60 days showed lower post-thaw viability compared to the ones thawed in the 60 days following cryopreservation (56.6±15.2% vs. 67.6±15.5%, p=0.04). Post-thaw graft viability did not influence engraftment time, but there was a predisposition towards infectious complications in the post-transplant period in patients receiving grafts with lower percentage of viable cells. They developed febrile neutropenia more often (72.2% vs. 50% of patients, p=0.05) and had more febrile days (2.4±2.6 vs. 1.5±2.3, p=0.05) following transplantation. We have demonstrated that PBPC grafts are capable of long term engraftment regardless of the graft storage time or percentage of viable cells post-thaw, which confirms the robustness of CD34+ cells during the freeze/thaw procedures carried out in daily clinical practice. Granulocyte concentration in PBPC grafts could have an influence on infectious complications following transplantation and needs to be further investigated on a larger number of patients

Post-thaw Viability of Cryopreserved Hematopoietic Progenitor Cell Grafts: Does It Matter?

Cell viability in peripheral blood progenitor cell (PBPC) grafts and its influence on the clinical course following transplantation was evaluated in 81 consecutive transplantations (72 autologous, 9 allogeneic) performed in patients with hematological diseases. Viability of cells in PBPC grafts immediately upon collection was 98.6±3.5%, after addition of dimethyl sulfoxide (DMSO) 73.3±21.8%, and post-thaw 65.2±16.1%. It did not differ significantly between patients with different diagnoses, gender, age, type of priming used, dose of G-CSF administered or number of CD34+ cells collected. However, grafts stored for more than 60 days showed lower post-thaw viability compared to the ones thawed in the 60 days following cryopreservation (56.6±15.2% vs. 67.6±15.5%, p=0.04). Post-thaw graft viability did not influence engraftment time, but there was a predisposition towards infectious complications in the post-transplant period in patients receiving grafts with lower percentage of viable cells. They developed febrile neutropenia more often (72.2% vs. 50% of patients, p=0.05) and had more febrile days (2.4±2.6 vs. 1.5±2.3, p=0.05) following transplantation. We have demonstrated that PBPC grafts are capable of long term engraftment regardless of the graft storage time or percentage of viable cells post-thaw, which confirms the robustness of CD34+ cells during the freeze/thaw procedures carried out in daily clinical practice. Granulocyte concentration in PBPC grafts could have an influence on infectious complications following transplantation and needs to be further investigated on a larger number of patients

THE ROLE OF NOTCH1 RECEPTOR, JAGGED1 AND DLL1 LIGAND EXPRESSION IN HEMATOPOIETIC STEM CELLS OF THE MICROENVIRONMENT IN MULTIPLE MYELOMA

Signalni put Notch u krvotvornim matičnim stanicama (KMS) koštane srži (KS) multiplog mijeloma (MM) analiziran je kroz ekspresiju proteina NOTCH1, JAGGED1, DLL1 u KMS imunofenotipizacijom, u tumorskim plazma stanicama (TPS) imunohistokemijskim bojenjem, uz analizu citogenetičkih promjena visokog rizika (CGVR) u TPS metodom FISH te analizu histoloških promjena u koštanoj srži MM. Ekspresija JAGGED1 i DLL1 u KMS značajno je niža u MM nego u zdravih darivatelja. Prisutna je negativna korelacija između ekspresije NOTCH1 i JAGGED1 u KMS MM, a pozitivna korelacija između ekspresije NOTCH1 i liganada JAGGED1 i DLL1 u TPS. U TPS s prisutnom barem jednom CGVR i difuznim infiltratom KS značajno je veća razina ekspresije NOTCH1 i JAGGED1 i intenziteta ekspresije NOTCH1. Potvrđena je hipoteza da postoji razlika u ekspresiji JAGGED1 i DLL1 u KMS u MM i zdravoj KS, no nije dokazana povezanost ekspresije NOTCH1, JAGGED1, DLL1 u KMS i TPS u MM. Povezanost ekspresije NOTCH1, JAGGED1 i DLL1 u istoj vrsti stanica ukazuje na razliku u homotipnoj i heterotipnoj staničnoj signalizaciji puta Notch u MM.The Notch signaling pathway in hematopoietic stem cells (HSC) in the multiple myeloma (MM) bone marrow (BM) was analyzed with the NOTCH1, JAGGED1, DLL1 expression in HSC by immunophenotyping and in tumor plasma cells (TPC) by immunohistochemical staining. High risk cytogenetic changes (CGHR) using the FISH method and histological analysis of TPC in the BM MM was performed. JAGGED1 and DLL1 expression in HSC were significantly lower in MM than in healthy donors. There was a negative correlation between NOTCH1 and JAGGED1 expression in HSC MM, and a positive correlation between NOTCH1 and JAGGED1 and DLL1 in TPC, respectively. In TPC with at least one CGHR present and a BM diffuse infiltrate, the NOTCH1 and JAGGED1 expression and the intensity of NOTCH1 expression were significantly higher. The hypothesis that there is a difference in the expression of JAGGED1 and DLL1 in HSC between MM and healthy BM was confirmed, but the correlation of the NOTCH1, JAGGED1, DLL1 expression in HSC and TPC in MM was not proven. The association of NOTCH1, JAGGED1 and DLL1 expression in the same cell type indicates a difference in homotypic and heterotypic cellular signaling of the Notch pathway in MM

THE ROLE OF NOTCH1 RECEPTOR, JAGGED1 AND DLL1 LIGAND EXPRESSION IN HEMATOPOIETIC STEM CELLS OF THE MICROENVIRONMENT IN MULTIPLE MYELOMA

Signalni put Notch u krvotvornim matičnim stanicama (KMS) koštane srži (KS) multiplog mijeloma (MM) analiziran je kroz ekspresiju proteina NOTCH1, JAGGED1, DLL1 u KMS imunofenotipizacijom, u tumorskim plazma stanicama (TPS) imunohistokemijskim bojenjem, uz analizu citogenetičkih promjena visokog rizika (CGVR) u TPS metodom FISH te analizu histoloških promjena u koštanoj srži MM. Ekspresija JAGGED1 i DLL1 u KMS značajno je niža u MM nego u zdravih darivatelja. Prisutna je negativna korelacija između ekspresije NOTCH1 i JAGGED1 u KMS MM, a pozitivna korelacija između ekspresije NOTCH1 i liganada JAGGED1 i DLL1 u TPS. U TPS s prisutnom barem jednom CGVR i difuznim infiltratom KS značajno je veća razina ekspresije NOTCH1 i JAGGED1 i intenziteta ekspresije NOTCH1. Potvrđena je hipoteza da postoji razlika u ekspresiji JAGGED1 i DLL1 u KMS u MM i zdravoj KS, no nije dokazana povezanost ekspresije NOTCH1, JAGGED1, DLL1 u KMS i TPS u MM. Povezanost ekspresije NOTCH1, JAGGED1 i DLL1 u istoj vrsti stanica ukazuje na razliku u homotipnoj i heterotipnoj staničnoj signalizaciji puta Notch u MM.The Notch signaling pathway in hematopoietic stem cells (HSC) in the multiple myeloma (MM) bone marrow (BM) was analyzed with the NOTCH1, JAGGED1, DLL1 expression in HSC by immunophenotyping and in tumor plasma cells (TPC) by immunohistochemical staining. High risk cytogenetic changes (CGHR) using the FISH method and histological analysis of TPC in the BM MM was performed. JAGGED1 and DLL1 expression in HSC were significantly lower in MM than in healthy donors. There was a negative correlation between NOTCH1 and JAGGED1 expression in HSC MM, and a positive correlation between NOTCH1 and JAGGED1 and DLL1 in TPC, respectively. In TPC with at least one CGHR present and a BM diffuse infiltrate, the NOTCH1 and JAGGED1 expression and the intensity of NOTCH1 expression were significantly higher. The hypothesis that there is a difference in the expression of JAGGED1 and DLL1 in HSC between MM and healthy BM was confirmed, but the correlation of the NOTCH1, JAGGED1, DLL1 expression in HSC and TPC in MM was not proven. The association of NOTCH1, JAGGED1 and DLL1 expression in the same cell type indicates a difference in homotypic and heterotypic cellular signaling of the Notch pathway in MM

THE ROLE OF NOTCH1 RECEPTOR, JAGGED1 AND DLL1 LIGAND EXPRESSION IN HEMATOPOIETIC STEM CELLS OF THE MICROENVIRONMENT IN MULTIPLE MYELOMA

Signalni put Notch u krvotvornim matičnim stanicama (KMS) koštane srži (KS) multiplog mijeloma (MM) analiziran je kroz ekspresiju proteina NOTCH1, JAGGED1, DLL1 u KMS imunofenotipizacijom, u tumorskim plazma stanicama (TPS) imunohistokemijskim bojenjem, uz analizu citogenetičkih promjena visokog rizika (CGVR) u TPS metodom FISH te analizu histoloških promjena u koštanoj srži MM. Ekspresija JAGGED1 i DLL1 u KMS značajno je niža u MM nego u zdravih darivatelja. Prisutna je negativna korelacija između ekspresije NOTCH1 i JAGGED1 u KMS MM, a pozitivna korelacija između ekspresije NOTCH1 i liganada JAGGED1 i DLL1 u TPS. U TPS s prisutnom barem jednom CGVR i difuznim infiltratom KS značajno je veća razina ekspresije NOTCH1 i JAGGED1 i intenziteta ekspresije NOTCH1. Potvrđena je hipoteza da postoji razlika u ekspresiji JAGGED1 i DLL1 u KMS u MM i zdravoj KS, no nije dokazana povezanost ekspresije NOTCH1, JAGGED1, DLL1 u KMS i TPS u MM. Povezanost ekspresije NOTCH1, JAGGED1 i DLL1 u istoj vrsti stanica ukazuje na razliku u homotipnoj i heterotipnoj staničnoj signalizaciji puta Notch u MM.The Notch signaling pathway in hematopoietic stem cells (HSC) in the multiple myeloma (MM) bone marrow (BM) was analyzed with the NOTCH1, JAGGED1, DLL1 expression in HSC by immunophenotyping and in tumor plasma cells (TPC) by immunohistochemical staining. High risk cytogenetic changes (CGHR) using the FISH method and histological analysis of TPC in the BM MM was performed. JAGGED1 and DLL1 expression in HSC were significantly lower in MM than in healthy donors. There was a negative correlation between NOTCH1 and JAGGED1 expression in HSC MM, and a positive correlation between NOTCH1 and JAGGED1 and DLL1 in TPC, respectively. In TPC with at least one CGHR present and a BM diffuse infiltrate, the NOTCH1 and JAGGED1 expression and the intensity of NOTCH1 expression were significantly higher. The hypothesis that there is a difference in the expression of JAGGED1 and DLL1 in HSC between MM and healthy BM was confirmed, but the correlation of the NOTCH1, JAGGED1, DLL1 expression in HSC and TPC in MM was not proven. The association of NOTCH1, JAGGED1 and DLL1 expression in the same cell type indicates a difference in homotypic and heterotypic cellular signaling of the Notch pathway in MM

Post-thaw viability of cryopreserved hematopoietic progenitor cell grafts: does it matter? [Vijabilnost pripravka krvotvornih matičnih stanica nakon odmrzavanja: ima li kliničko značenje ili ne?]

Cell viability in peripheral blood progenitor cell (PBPC) grafts and its influence on the clinical course following transplantation was evaluated in 81 consecutive transplantations (72 autologous, 9 allogeneic) performed in patients with hematological diseases. Viability of cells in PBPC grafts immediately upon collection was 98.6±3.5%, after addition of dimethyl sulfoxide (DMSO) 73.3±21.8%, and post-thaw 65.2±16.1%. It did not differ significantly between patients with different diagnoses, gender, age, type of priming used, dose of G-CSF administered or number of CD34+ cells collected. However, grafts stored for more than 60 days showed lower post-thaw viability compared to the ones thawed in the 60 days following cryopreservation (56.6±15.2% vs. 67.6±15.5%, p=0.04). Post-thaw graft viability did not influence engraftment time, but there was a predisposition towards infectious complications in the post-transplant period in patients receiving grafts with lower percentage of viable cells. They developed febrile neutropenia more often (72.2% vs. 50% of patients, p=0.05) and had more febrile days (2.4±2.6 vs. 1.5±2.3, p=0.05) following transplantation. We have demonstrated that PBPC grafts are capable of long term engraftment regardless of the graft storage time or percentage of viable cells post-thaw, which confirms the robustness of CD34+ cells during the freeze/thaw procedures carried out in daily clinical practice. Granulocyte concentration in PBPC grafts could have an influence on infectious complications following transplantation and needs to be further investigated on a larger number of patients