Non-cryopreserved Peripheral Blood Stem Cells Research Articles

The use of non-cryopreserved hematopoietic stem cells for autologous transplantation in multiple myeloma patients

Background . An autologous stem cell transplant (ASCT) is the standard of treatment young and fit patients with multiple myeloma (MM). The using this method of treatment is limited due to the high consumption of economic and intellectual resources, the low availability of Cryobank departments in regions of Russia. Objective : to evaluate the efficacy and safety of using non-cryopreserved peripheral blood stem cells (non-CRYO PBSCs) in patients with multiple myeloma who underwent autologous transplantation. Materials and methods . We conducted a prospective study of the efficacy and safety of using non-CRYO PBSCs in patients with MM who underwent ASCT. The number of PBSCs apheresis procedures in 82.9 % (n = 29 pts) was 1 day, and in 17.1 % (n = 6 pts) was 2 days. After apheresis, PBSCs were stored at a temperature of +4…+6 °C in a blood bank refrigerator for up to 72 hours. During 4 years, ASCT using non-CRYO PBSCs was performed in 35 patients with MM. The average number CD34+ cell dose was 2.63 × 106 /kg. The median time to neutrophil engraftment was 11 days (range from 9 to 14). The median time to platelet engraftment was 12 days (range from 8 to 19). The control group included 43 patients with MM, who underwent PBSCs ASCT according to the traditional method, including preparation and cryopreservation of the apheresis product of PBSCs with cryoprotectant dimethyl sulfoxide, as well as subsequent defrosting on the day of transplantation (day 0). Results . There were no significant differences comparing the safety of ASCT in non-CRYO and CRYO PBSCs groups among such parameters as the viability of PBSCs, the frequency of complications, the time of hematopoietic engraftment, the need for platelet and red blood cells transfusion therapy, and time of hospitalization. Conclusion . Hence, the method of short-term storaging non-CRYO PBSCs is not inferior to the traditional method of controlled freezing, is more economical and can be used in medical organizations that do not have a Cryobank in their structure.

NON Cryopreserved Hematopoietic STEM CELLS to Autograft Patients with Lymphomas: A PAIR Matched Analysis Comparing a Single Center Experience to the Use of Cryopreserved STEM CELLS Reported to the EBMT Registry

Introduction: Around fifty thousand autologous stem cell transplantations are done each year worldwide, using cryopreserved peripheral blood stem cells (PBSC). Cryopreservation is time consuming and expensive. Since 2007, several retrospective studies have shown that PBSC can be stored at 4Dg C for two to three days, allowing autologous stem cell transplantation (ASCT) in patients with multiple myeloma receiving high dose melphalan. Data with non-cryopreserved PBSC in patients autografted for lymphoma following longer preconditioning regimens are limited. In addition, there has been no controlled comparison able to possibly detect unforeseen differences. Patients and methods: We compared outcomes of 94 consecutive adult patients with lymphomas (66 with Hodgkin Lymphoma) autografted in our department in Oran (Algeria), using PBSC stored at 4 dg C, from 2009 to 2018 with patients receiving cryopreserved stem cells reported to the EBMT registry. Patient's autografted in Oran were matched with patients receiving cryopreserved PBSC in the registry (4 controls per Oran Patient). Results: Neutrophil engraftment was significantly faster with cryopreserved PBSC (p= 0.003): by day 10, only 17% of patients receiving non-cryopreserved PBSC engrafted versus 48% for cryopreserved PBSC. Likewise, platelet recovery to 20 000/mm3 was significantly faster in patients receiving cryopreserved PBSC (p=0.01). However, all patients in both groups had recovered by day 20. There were no significant differences in Non Relapse Mortality (9% versus 7%; p=0.4), Relapse Incidence (22% versus 32%; p=0.13), Progression Free Survival (70% versus 61%; 0.4) or Overall Survival (85% versus 75%; p=0.3). Conclusion: This analysis suggests that, in patients with lymphomas receiving pretransplant regimens such as the BEAM, PBSC stored at 4dg C for up to six days can be used safely in centers with no cryopreservation facility. The kinetics of recovery of hematopoiesis however did show a significant, albeit small, delay of engraftment for both neutrophils and platelets, which favors the use of cryopreservation if available. Disclosures Blaise: Jazz Pharmaceuticals: Honoraria. Peggs:Autolus: Consultancy. Salles:Karyopharm: Consultancy; F. Hoffman-La Roche Ltd: Consultancy, Honoraria, Other; Bristol Myers Squibb: Consultancy, Other; Celgene: Consultancy, Honoraria, Other: Participation in educational events; Autolus: Consultancy; Abbvie: Consultancy, Honoraria, Other: Participation in educational events; Genmab: Consultancy; Novartis: Consultancy, Honoraria, Other; Kite: Consultancy, Honoraria, Other; Takeda: Consultancy, Honoraria, Other; Debiopharm: Consultancy; Amgen: Honoraria, Other: Participation in educational events; MorphoSys: Consultancy, Honoraria, Other; Gilead: Consultancy, Honoraria, Other: Participation in educational events; Janssen: Consultancy, Honoraria, Other: Participation in educational events; Epizyme: Consultancy. Milpied:Roche: Honoraria, Other: Travel support; Astellas: Honoraria; Sandoz: Honoraria, Other: consultancy or advisory role; Janssen: Honoraria; Gilead Sciences: Other: consultancy or advisory role; Celgene: Other: Travel support.

Non-cryopreserved peripheral blood stem cells autologous transplantation in multiple myeloma: Bicentric study.

The objective of this study is to evaluate the toxicity of autologous transplantation of non-frozen peripheral blood stem cells in Moroccan patients with multiple myeloma. This was a bicentric retrospective study conducted in the Clinical Haematology Department of Mohammed V Military Teaching Hospital and at the Al Madina Clinic in Casablanca. The study period was from January 2015 to June 2019. All patients with multiple myeloma who had undergone an autologous peripheral stem cell transplant without freezing were included. Mobilisation was performed with lenograstim alone and the collected stem cells were stored for 24-48hours in a blood bank refrigerator at a temperature of 4°C. After standard conditioning with high-dose melphalan, the peripheral blood stem cells were reinjected 24h following conditioning. Over the study period, 55 patients received an autologous transplant using non-frozen peripheral blood stem cells. The median richness of the CD34 cells collected was 4.5×106 CD34/kg (range: 2-12.2). The time required for neutrophil recovery was 12 days (range: 7-19). The time required for platelet recovery was 14 days (range: 9-32). The mortality rate within 100 days post-transplant was 3.6%. We did not observe any cases of graft failure. Our study finds good feasibility and low toxicity of autologous peripheral stem cell transplantation without freezing in patients with multiple myeloma.

Short-Term Noncryopreserved Hematopoietic Stem Cells: Single Center Experience of Autologous Transplantation

Background: High-dose chemotherapy (HDCT) followed by autologous transplantation of hematopoietic stem cells (auto-HSCT) is an important component of treatment in multiple myeloma (MM). There is a standard method of controlled cryopreservation of HSC suspension. We found that the storage of native HSC suspension with temperature fluctuations from +3 °C to +5 °C during 72 - 120 hours does not significantly affect the content of CD34+ cells in the product, the index 7AAD- (7-AAD (7-aminoactinomycin - D) is a fluorescent marker that penetrates damaged cell membranes and binds to double-stranded DNA. Through 7AAD does not penetrate intact membranes, so living cells are not stained 7AAD with flow cytometry), and colony-forming ability (CFA) of HSC, as well as the recovery time of hematopoiesis in MM patients after auto-HSCT. Aim: To evaluate the effectiveness and safety of the method of storage of non-cryopreserved peripheral blood stem cells. Methods: 39 patients with MM were included in this study(male/female ratio 1.36:1). All the patients get standard immunochemotherapy programs and were in remission at the time of auto-HSCT. Patients were divided into two groups depending on the method of stem cell storage: group 1 - non-cryopreserved (n=20), group 2 - cryopreserved (standard) (n=19). An effectivity and safety were evaluated in such parameters as the number of CD34+ and 7AAD- cells, CFA after apheresis and before reinfusion of HSC. Also, we evaluated the number of platelets concentrate transfusions, the timing of engraftment of granulocytic and megakaryocytic blood sprouts, the length of hospital stays after auto-HSCT. Results: The results are presented in the comparison table of the evaluated parameters. Our data showed significantly reduce of episodes febrile neutropenia and cases of enteropathy. Conclusion: Thus, the proposed method of storage of HSC is not inferior to the traditional method with cryopreservation on such parameters as CD34+, 7AAD-, CFA, the number of platelets concentrate transfusions, terms of hematopoiesis restoration, length of hospital stay after HSCT, the number of complications. Table. Disclosures Shuvaev: Fusion Pharma: Consultancy; BMS: Consultancy; Novartis: Consultancy; Pfize: Honoraria.

Use of Non-Cryopreserved Peripheral Blood Stem Cells Is Associated with Adequate Engraftment in Patients with Multiple Myeloma Undergoing an Autologous Transplant

Autologous transplantation is the standard of care for transplant-eligible patients with multiple myeloma. Toward making this treatment accessible in developing countries, there are significant challenges like resource constraints and access to cryopreservation facilities. We performed a retrospective analysis of patients with multiple myeloma who underwent autologous transplantation using granulocyte colony-stimulating factor (G-CSF)-mobilized non-cryopreserved grafts at our institution from January 1995 to December 2014. Peripheral blood stem cells (PBSCs) were harvested over 1 to 2 days after G-CSF mobilization. After apheresis, PBSCs were stored at 4°C in a blood bank refrigerator for up to 72 hours. During the study period, 224 patients with multiple myeloma underwent autologous transplantation using G-CSF–mobilized non-cryopreserved grafts. The number of days of stem cell harvest was 1 in 91 patients (40.6%) and 2 in 133 patients (59.4%). The median CD34 cell dose was 4.87 × 106/kg (range, 1.15 to 23.7). All patients except 1 engrafted. The median time to neutrophil engraftment was 12 days (range, 9 to 22). The median time to platelet engraftment was 17 days (range, 10 to 44). In a resource-limited setting, the use of G-CSF–mobilized non-cryopreserved grafts results in adequate engraftment for most patients with multiple myeloma undergoing autologous stem cell transplantation.

A Feasibility Study of the Full Outpatient Conduction of Hematopoietic Transplants in Persons with Multiple Sclerosis Employing Autologous Non-Cryopreserved Peripheral Blood Stem Cells

Background: With the goal of achieving immune system reset, autologous hematopoietic stem cell transplantations have been performed in patients with multiple sclerosis (MS). Material and Methods: Two hundred and eighty-six consecutive patients with MS were autografted in a single center using non-frozen peripheral blood stem cells (PBSCs), on an outpatient basis and conditioning with cyclophosphamide and rituximab. The protocol was registered in ClinicalTrials.gov identifier NCT02674217. Results: One hundred and ninety-four females and 92 males were included; the median age was 47. All procedures were started on an outpatient basis and only 8 persons needed to be admitted to the hospital during the procedure. In order to obtain at least 1 × 10⁶/kg viable CD34 cells, 1-4 aphereses were performed (median 1). The total number of viable CD34+ cells infused ranged between 1 and 19.2 × 10⁶/kg (median 4.6). Patients recovered above 0.5 × 10⁹/L absolute granulocytes on median day 8 (range 0-12). Two individuals needed red blood cells but none needed platelet transfusions. There were no transplant-related deaths and the 128-month overall survival of the patients is 100%. In 82 persons followed up for 3 or more months, the Expanded Disability Status Scale diminished from a mean of 5.2-4.9, the best results being obtained in relapsing-remitting and primary progressive MS. Conclusions: It is possible to conduct autotransplants for patients with MS employing non-frozen PBSCs and outpatient conduction. Additional information is needed to assess the efficacy of these procedures in the treatment of patients with MS.

159 - A Feasibility Study of the Full Outpatient Conduction of Hematopoietic Transplants in Persons with Multiple Sclerosis Employing Autologous Non-Cryopreserved Peripheral Blood Stem Cells

159 - A Feasibility Study of the Full Outpatient Conduction of Hematopoietic Transplants in Persons with Multiple Sclerosis Employing Autologous Non-Cryopreserved Peripheral Blood Stem Cells

A Feasibility Study of the Full Outpatient Conduction of Hematopoietic Transplants in Persons with Multiple Sclerosis Employing Autologous Non-Cryopreserved Peripheral Blood Stem Cells

Background: With the goal of immune system reset, autologous hematopoietic stem cell transplantations have been done in patients with multiple sclerosis (MS).Material and methods: 131 consecutive patients with MS were autografted in a single center using non-frozen peripheral blood stem cells (PBSC) on an outpatient basis and conditioning with cyclophosphamide (Cy) and rituximab. The protocol was registered in ClinicalTrials.gov identifier NCT02674217. The PBSC mobilization schedule was done with Cy and filgrastim (G-CSF). Intravenous Cy (50 mg/Kg) was delivered on days - 11 and - 10. Subcutaneous G-CSF (10 ug / Kg / bid) was delivered on days - 9 to - 1.. The apheresis procedure was performed on day - 2. The apheresis objective was to reach at least 1 x 106 viable CD34+ cells/Kg. As outpatients and after collecting the PBSC, intravenous Cy (50 mg / Kg) was delivered along a 120 minute period, on days - 2 and - 1 followed by MESNA (1000 mg/m2 along a 180-minute period). After the intravenous Cy, oral ondansetron), oral cotrimoxazole, oral fluconazole and oral acyclovir were used in all patients until granulocytes were greater than 0.5 x 109/L. After the recovery of the granulocytes, patients were given rituximab (375 mg/m2 along a 3 h period) and subsequently rituximab (100 mg) every two months along a 12-month period. The cumulative dose of Cy is 200 mg/Kg.Results: 80 females and 51 males were included; median age was 47 years. All procedures were started on an outpatient basis and two persons were admitted to the hospital during the procedure. In order to obtain at least 1 x106 / Kg viable CD34 cells, one to four apheresis were performed (median 1). Total number of viable CD34+ cells infused ranged between 1 and 9.6 x106 / Kg (median 2.2). Patients recovered above 0.5 x109/L absolute granulocytes on median day 9 (range 6 to 12). Two individuals needed red blood cells but none needed platelet transfusions. There were no transplant related deaths, the 120-month overall survival being 100%. In a subset of 25 persons followed for 5 months or more the EDSS was assessed three months after the graft and means diminished from 5.4 to 4.9. The EDSS score improved in 11 patients (44%), remained stable in 7 (28%) and worsened in 7 (28%). A tendency to diminish the EDSS score as a function of time after the autograft was observed in this subset of individuals.Conclusions: It is possible to conduct autotrasplants for patients with MS employing non-frozen peripheral blood stem cells and outpatient conduction. Additional information is needed to assess the efficacy of these procedures in the treatment of patients with MS. DisclosuresNo relevant conflicts of interest to declare.

Outpatient Hematopoietic Grafting in Patients with Multiple Sclerosis Employing Autologous Non-Cryopreserved Peripheral Blood Stem Cells: A Feasibility Study

Background:Multiple sclerosis (MS) is a chronic, inflammatory, debilitating disease that causes destruction of central nervous system (CNS) myelin, with varying degrees of axonal damage. With the goalofd re-setting the immune system, autologous hematopoietic stem cell transplantations (HSCT) have been done in patients with MS since 1996 and more than 700 HSCTs have been performed around the world. The risk of transplant related mortality in HSCT for MS has declined over the past years.Material and methods:Consecutive patients with MS were autografted in a single center using: Hematopoietic stem cells (HSC) were mobilized with cyclophosphamide (Cy), 3 gr/m2 and G-CSF, the procedure was conducted on outpatient basis employing peripheral blood non-frozen HSC and conditioning with high-dose Cy (100 mg/Kg) and post-transplant G-CSF and rituximab. Antibiotics, antimycotics and antivirals were given orally.Results:Thirteen patients with MS were prospectively accrued in the study. There were 7 females and 6 males. Median age was 48 years, range 24 to 65. The expanded disability status scale (EDSS) score of these patients had a median of 5 points (range 1 to 6). All the autografts were started on an outpatient basis and two persons were admitted to the hospital during the procedure (persistent nausea/vomiting and neutropenic fever); they stayed in the hospital for 48 hours. In order to obtain a minimum of 1 x106 viable CD34+ cells/Kg, one to four apheresis were done (median 1). The total number of viable CD34+ cells infused to the patients ranged between 1 and 9.6x106 (median 3.1). Patients recovered above 0.5 x109/L absolute granulocytes on median day 9 (range 6 to 12). No individuals needed transfusions of red blood cells nor platelets transfusions. There were no transplant-related deaths and the 23-month overall survival of the autografted patients is 100%. Median cost of the procedure was 30 000 USD. In 8 persons the EDSS was assessed three months after the graft; it diminished from a median of 4.5 to a median of 2.5. In 5 patients, the three months re-assessment of the EDSS has not been possible as a result of the time elapsed after the autograft.Discussion:These data indicate that it is possible to conduct autotrasplants for patients with MS employing a simplification of the conventional procedures by means of non-frozen peripheral blood stem cells and outpatient conduction. Additional information is needed to asses the efficacy of these procedures in the treatment of patients with MS. DisclosuresNo relevant conflicts of interest to declare.

Cell-Freezing Devices Are Not Strictly Needed to Start an Autologous Hematopoietic Transplantation Program: Non-Cryopreserved Peripheral Blood Stem Cells Can be Used to Restore Hematopoiesis after High Dose Chemotherapy: A Multicenter Experience in 268 Autografts in Patients with Multiple Myeloma or Lymphoma. Study on Behalf of the Latin-American Bone Marrow Transplantation Group (LABMT)

Storage of peripheral blood stem cells (PBSC) at 4ºC is a simple and inexpensive alternative to cryopreservation for preserving the clonogenic capacity of progenitors cells in the autologous transplant setting, however it has been perceived as unsafe and has deserved little attention. We present the experience of two Latin-American centers using refrigerated, non-cryopreserved stem cells after conditioning with high dose melphalan, CBV or BEAM in a large group of lymphoma and myeloma patientsMaterials and MethodsPBSC were mobilized with filgrastim 5 mg/kg/BID for three to six days.One to three apheresis procedure were employed; the cells were stored at 4ºC for 5 to 6 days in patients who received BEAM or CBV and for 3 days in those given melphalan. All of the conditioning regimens were administered preserving the full intensity of dose (Table 1). After the autograft all patients received filgrastim or pegfilgrastimTable 1BEAMD-5D-4D-3D-2D-1BiCNU 300 mgs/m2XEtoposide 200-400 mgs/m2XXXXCitarabine 300-400 mgs/m2XXXMelphalan 140 mgs/m2XCBVBiCNU 300 mgs/m2 13 patients received carboplatin 900 mgs/m2 instead BiCNUXEtoposide 300 mgs/m2XXXCiclophosphamide 2.000 mgs/m2XXXMelphalanMelphalan 200 mgs/m2XMelphalan 100 mgs/m2XXResults102 lymphoma patients: 48 Hodgkin's lymphoma (HL) and 54 non-Hodgkin´s lymphoma (NHL) received BEAM (71) or CBV (31). A median of 3.3 millions/kg of CD34 was infused; the median viability of the cells after 5-6 days of refrigeration (trypan blue exclusion) was 82%. 101 out of 102 patients engrafted, median time to achieve 500/ul neutrophil or more was 12 days, 100 were evaluable for thrombopoiesis, 99 of them had a self- sustained platelet count of 20.000 or more at a median of 17 days. The OS at 5 years was 59% and 42% for patients with Hodgkin and lymphoma respectively151 patients with multiple myeloma received melphalan 200 mgs/m2. After 72 hours of refrigeration, a median of 2.6 millions/kg of CD34 cells were infused, the viability in all cases being > 90%. Three patients were not evaluable because early death. Median time to achieve 500 neutrophil or more and 20.000 platelets was 12 (9-50) and 15 (7-50) days. The OS at 5 years was 50%21 patients with NHL and HL received as conditioning regimen melphalan 200 mgs/m2. After 72 hours of storage, a median of 1.75 millions/kg of CD34 cells were transplanted, 100% of them engrafted, median time to 500 neutrophils and 20.000 platelets was 11.9 and 15 days respectivelyThere were no cases of secondary engraftment failure in any of the three groupsConclusionIn this series of 268 patients, we have shown that autologous PBSC can be kept at 4ºC in a conventional blood bank refrigerator for up to six days and use them to rescue high-dose chemotherapy in both multiple myeloma and lymphoma patients.Avoiding freezing procedures results in substantial cost savings. The availability of freezing devices for hematopoietic stem cells is not anymore an obstacle to start a an autologous transplantation program This observation is critical in areas of underprivileged economic circumstances, where more than 50% of the inhabitants of the world live. DisclosuresNo relevant conflicts of interest to declare.

High-Dose Chemotherapy and Autologous Stem Cell Transplantation in Multiple Myeloma: A Single Institution Experience at All India Institute of Medical Sciences, New Delhi, Using Non-Cryopreserved Peripheral Blood Stem Cells

Intravenous high-dose melphalan has a short half-life, and application of this single drug in MM transplant favors the use of stem cells without cryopreservation, for wider use in general and in resource-limited settings in particular. Ninety-two patients with MM were given high-dose melphalan and rescued with granulocyte colony stimulating factor (G-CSF) mobilized noncryopreserved autologous PBSC, in our hospital during the past 18 years. Stem cells were mobilized with 4 days of G-CSF, harvested (median CD34 dose, 2.9 × 10(6)/kg) and then stored at 4°C in a refrigerator for a median of 2 days (range, 1-5 days) before reinfusion. Median time to neutrophil (> 500/mm(3)) and platelet (> 20,000/mm(3)) engraftment were 10 and 14 days respectively. There was no graft failure. Mucositis grade 3/4 was seen in 66 patients (72%). Transplant-related mortality at 100 days was 3.2%. The overall response to transplant was 88% and improvement compared with pretransplant status was seen in 48%. The median overall survival (OS) and progression-free survival (PFS) were 61.7 months and 35.4 months respectively; independent predictors of survival were Eastern Cooperative Oncology Group Performance Status and hemoglobin for OS and chemosensitive disease and remission status after transplant for PFS. We conclude that high-dose chemotherapy and autologous transplant with noncryopreserved PBSC is a simple, effective, and safe method for MM with equivalent results, and that cryopreservation is not necessary. It reduces the cost of transplant and avoids dimethyl sulfoxide toxicity.

High-Dose Chemotherapy Plus Non-Cryopreserved Autologous Peripheral Blood Stem Cell Transplantation Rescue for Patients With Refractory or Relapsed Hodgkin Disease

A simplified schedule of high-dose chemotherapy consisting of cyclophosphamide (60 mg · kg −1 · d −1 for 2 days), etoposide (15 mg · kg −1 · d −1 for 2 days), and carboplatin (400 mg/m 2 per day for 2 days) plus autologous non-cryopreserved peripheral blood stem cells (PBSCs) was used for treatment of patients with relapsed (n = 25) and refractory (n = 3) Hodgkin disease. The use of such PBSCs mobilized by granulocyte colony-stimulating factor after high-dose myeloablative therapy resulted in a rapid, complete, and sustained hematopoietic recovery. The median time to achieve an absolute neutrophil count >0.5 × 10 9/L was 13 days (range, 7-18 days). The median time to a self-sustained platelet count >20 × 10 9/L was 15 days (range, 7-20 days). Twelve of the 28 patients (43%) were alive and without disease at a median follow-up of 16 months (range, 9-86 months) for all surviving patients. The estimated 2-year overall survival and disease-free survival for all patients were 45% and 42%, respectively. Thirteen patients died of relapse or progressive disease, 2 died of infection, and 1 was still surviving in relapse by the time of the analysis. The median time to relapse was 10 months (range, 3-28 months) from PBSC infusion. High-dose chemotherapy with short-duration chemotherapy and non-cryopreserved bone marrow is an effective and safe treatment modality for patients with relapsed or resistant Hodgkin lymphoma.

Cyclophosphamide, etoposide and carboplatine plus non-cryopreserved autologous peripheral blood stem cell transplantation rescue for patients with refractory or relapsed non-Hodgkin's lymphomas

A simplified schedule of high-dose chemotherapy consisting of cyclophosphamide (60 mg/kg/day for 2 days), etoposide (15 mg/kg/day for 2 days) and carboplatine (400 mg/m(2)/day for 2 days), together with autologous non-cryopreserved peripheral blood stem cells was used for treatment of relapsed (29 patients) and refractory (three patients) patients with non-Hodgkin's lymphoma (NHL). The use of such granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cells (PBSC) after high-dose myeloablative therapy resulted in a rapid, complete and sustained hematopoietic recovery. The median time to achieve an absolute neutrophil count greater than 0.5 x 10(9)/l was 12 days (range 8-17 days). The median time to self-sustained platelet count greater than 20 x 10(9)/l was 14 days (range 7-19 days). Fifteen of the 32 patients (49%) were alive and disease free at a median follow-up of 18 months (range 10-96 months) for all surviving patients. The estimated 2-year overall survival (OS) and disease free survival (DFS) for all patients were 50 and 43%, respectively. Twelve patients died of relapse or progressive disease, two patients died of infection and one patient died of cardiac cause. The median time to relapse was 12 months (5-27) from PBSC infusion. High-dose chemotherapy with short-duration chemotherapy and non-cryopreserved bone marrow (BM) is an effective and safe treatment modality for patients with relapsed or resistant NHL.