Recent clinical data, including a large scale multi-center study, suggest that there will be a continued role for harvested bone marrow as a source of hematopoietic progenitor cells (HPC) for allogeneic transplantation. As there is a significant erythrocyte content in harvested bone marrow, major ABO mismatched transplants generally require some form of erythrocyte depletion from the graft to prevent potential morbidity or mortality from hemolytic reactions at infusion. We have previously reported adaptation of the Haemonetics CellSaver™ for laboratory use for Ficoll separation of MNC. We have now added simple buffy coat formation for the purpose of erythrocyte depletion to our repertoire of functions for this instrument which is primarily marketed for intra-operative blood salvage. Unprocessed marrow is separated in a bell-shaped bowl that is spinning at 4800rpm. Marrow is pumped into the bowl at 100 mL/min until the bowl is approximately ¾ full at which point pump speed is reduced to 20 mL/min. A buffy coat layer is observed moving to the top of the packed RBC content of the bowl. As the buffy coat layer reaches the top of the bowl, the waste bag clamps are closed, and the collection bag clamps are opened to effect collection. Once the buffy coat layer has been collected, the pump and bowl centrifuge are stopped. The clamps are then closed on the collection bag, and reopened on the waste bag. The bowl is then emptied of the buffy coat depleted marrow. The red blood cells from the depleted marrow are saved in a satellite bag to be used in further processing of the marrow when additional volume is needed to fill the bowl during a sequence. The sequence above is repeated until the entire volume of marrow has been processed. Depending on the initial volume of marrow to be processed, 1-4 sequences are performed. We have performed this procedure with two bowl sizes, namely 125 mL capacity (n=7) and 225 mL capacity (n=5). The extent of RBC depletion, and recovery of both TNC and CD34+ cells is reported in the table. Tabled 1 Bowl Size Mean RBC depletion % Mean TNC recovery % Mean CD34 recovery % 125 mL 97.7 55.3 65.1 225 mL 79.1 52.7 63.3 Open table in a new tab

Over the immediate past 4 years, our program has collected hematopoietic progenitor cells by apheresis from 48 individuals aged 61 and over (range 61–71 years of age). We have retrospectively analyzed the collection and transplant results associated with employing these donors, and have compared them with 175 donors aged 60 or less who were collected during the same time period. We have found no significant difference in venous access (P=0.208), rate of post-transplant engraftment of neutrophils (P=0.117) and platelets (P=0.692), or in rate and grade of acute GVHD (P=0.806). However, we have found that these older donors have a significantly lower mobilization of CD34+ cells as reflected in lower absolute counts of circulating CD34+ cells pre-apheresis (P=0.016). This, in turn, results in lower CD34+ cell yields in apheresis products (P<0.001), trending towards requiring more apheresis procedures (22.9 vs 13.7%, P=0.095) to collect sufficient CD34+ cells for transplantation. We conclude that it is practical when necessary to employ donors aged 60 and above, as well as safe for both donor and intended recipient. However, concern over reduced CD34+ cell mobilization may be sufficient grounds to seek younger donors when possible.

Abstract Abstract 4226 There has been reluctance to collect apheresis HPC grafts from donors of age 60 and beyond. This is predicated on concern over relative frailty, poor venous access, reduced potential for HPC mobilization, and reduced potential for stable engraftment of HPC transplant. We have expanded our transplant eligibility into patients beyond the age of 60, with the concomitant acceptance of matched sibling donors of similar age. We have reviewed 96 consecutive sibling donors, 19 of whom were of age ≥60 years at the time of collection. All donors were collected using Gambro Spectra apheresis instruments, and the volume of blood pheresed was gauged to target a CD34 dose of 5-10 million per Kg of recipient weight. We have found the following: Age < 60 (n=77) Age ≥ 60 (n=19) p Blood volumes (Mean±Std) 4.9±3.3 6.1±3.1 .85 CD34/RecipKg E6 (Mean±Std) 8.1±2.8 5.8±1.7 .02 CD34/Blood vol E7. (Mean±Std) 18.0±11.7 10.9±7.8 .06 < 5E6 CD34/Kg Collected (n) 5 3 .21 > 1 collection (n) 12 7 .04 Catheter required (n) 23 6 .54 Grade 3+ pheresis complications 2 3 .47 ANC500 Median days (Min-Max) 16 (10-28) 16 (13-75) .14 Plt20K Median days (Min-Max) 16 (8-77) 16 (13-89) .30 These results suggest that donors of age ≥60 years may be successfully collected, and that the resultant grafts can be expected to produce successful transplants. There is not a higher toxicity rate or need for catheter insertion associated with the collections in the older age group. There is a clear trend, however, to reduced mobilization of CD34+ cells, as reflected in the need for more collections, and fewer CD34+ cells per blood volume leukopheresed. In spite of this, sufficient cells to produce functional grafts were collected from all donors, although a limited number of donors in both age categories failed to collect a full 5e+6 CD34/Kg. We conclude that in the context of an aging demographic, allograft donors of age ≥60 may be successfully employed for HPC collection. Further, we propose that the application of plerixafor in ≥60 year old donors should be investigated in the context of a growing need to collect allografts from donors in this age group. Disclosures: Off Label Use: sirolimus for graft-versus-host disease.

Journal of HematotherapyVol. 4, No. 3 A Prospective Randomized Trial Comparing Blood- and Marrow-Derived Stem Cells for Hematopoietic Replacement Following High-Dose ChemotherapyWilliam E. Janssen, Renee C. Smilee, and Gerald J. ElfenbeinWilliam E. JanssenSearch for more papers by this author, Renee C. SmileeSearch for more papers by this author, and Gerald J. ElfenbeinSearch for more papers by this authorPublished Online:27 Mar 2009https://doi.org/10.1089/scd.1.1995.4.139AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail "A Prospective Randomized Trial Comparing Blood- and Marrow-Derived Stem Cells for Hematopoietic Replacement Following High-Dose Chemotherapy." , 4(3), pp. 139–140FiguresReferencesRelatedDetailsCited byA randomized controlled clinical trial to determine the optimum duration of G-CSF priming prior to BM stem cell harvestingCytotherapy, Vol. 9, No. 2Recent advances in allogeneic hematopoietic stem-cell transplantationJournal of Laboratory and Clinical Medicine, Vol. 141, No. 1Stem Cell MobilizationHematology, Vol. 2003, No. 1Blood Stem Cell Versus Bone Marrow Transplantation10 June 2013Immune dysfunction despite high levels of immunoregulatory cytokine gene expression in autologous peripheral blood stem cell transplanted non-Hodgkin's lymphoma patientsExperimental Hematology, Vol. 28, No. 5Characterization and Outcome of “Hard to Mobilize” Lymphoma Patients Undergoing Autologous Stem Cell Transplantation5 August 2009 | Leukemia & Lymphoma, Vol. 39, No. 5-6Does Recombinant Human Granulocyte Colony-Stimulating Factor Really Prime Marrow Stem Cells in Mice and Humans?Blood, Vol. 93, No. 12Clinical Factors Contributing to the Pace of Engraftment After Allogeneic and Autologous Stem Cell TransplantationStem Cell Collection for Hematopoietic Transplantation: Stem Cell Sources, Mobilization Strategies, and Factors that Influence YieldHemopoietic stem-cell harvesting and transplantation using G-CSF-primed BM: comparison with unprimed BM and G-CSF-primed PBSCCytotherapy, Vol. 1, No. 5Granulocyte-Colony Stimulating Factor (G-CSF)-Primed, Delayed Marrow Harvests as a Source of Hematopoietic Stem and Progenitor Cells for Allogeneic Transplantation1 July 2009 | Leukemia & Lymphoma, Vol. 35, No. 3-4Composition and function of peripheral blood stem and progenitor cell harvests from patients with severe active rheumatoid arthritis4 January 2002 | British Journal of Haematology, Vol. 103, No. 3Myeloid growth factors in oncology23 February 2005 | Expert Opinion on Investigational Drugs, Vol. 7, No. 12Comparison of rhG-CSF primed bone marrow and blood stem cell autografts: an analysis of engraftment in malignant lymphomas and solid tumours24 April 2009 | European Journal of Haematology, Vol. 61, No. 4In-vitro- und In-vivo-Manipulation hämatopoetischer Stammzellen für die HochdosischemotherapieConsensus conference on high-dose therapy with hematopoietic stem cell transplantation in diffuse large-cell lymphomaAnnals of Oncology, Vol. 9Clinical Trial: Hematopoietic Progenitor Cell Transplantation in Breast Cancer: Current Status and Future Directions11 June 2009 | Cancer Investigation, Vol. 16, No. 2The True Cost of Bone Marrow TransplantationThe American Journal of the Medical Sciences, Vol. 314, No. 2Randomized Trial of Autologous Filgrastim-Primed Bone Marrow Transplantation Versus Filgrastim-Mobilized Peripheral Blood Stem Cell Transplantation in Lymphoma PatientsBlood, Vol. 90, No. 1Hematopoietic transplantation: State of the artStem Cells, Vol. 15, No. S2The Biology and Clinical Uses of Blood Stem CellsBlood, Vol. 89, No. 7Potential use of hematopoietic stem cells after radiation injury4 June 2009 | STEM CELLS, Vol. 15, No. S1HOST IMMUNOLOGIC AUGMENTATION FOR THE CONTROL OF INFECTIONInfectious Disease Clinics of North America, Vol. 10, No. 2G-CSF-primed bone marrow cells for autologous transplantationThe Lancet, Vol. 347, No. 9008Optimizing recovery from aplasia after high-dose therapy and hematopoietic stem cell transplantation1 March 1996 | Journal of Oncology Pharmacy Practice, Vol. 2, No. 1_suppl Volume 4Issue 3Jun 1995 To cite this article:William E. Janssen, Renee C. Smilee, and Gerald J. Elfenbein.A Prospective Randomized Trial Comparing Blood- and Marrow-Derived Stem Cells for Hematopoietic Replacement Following High-Dose Chemotherapy.Journal of Hematotherapy.Jun 1995.139-140.http://doi.org/10.1089/scd.1.1995.4.139Published in Volume: 4 Issue 3: March 27, 2009PDF download

Bone Marrow Transplant Program, University of South Florida College of Medicine, Moffitt Cancer Center, Tampa, FL, USA

Journal of HematotherapyVol. 2, No. 1 When, How, What, and Where to Purge Are Not the Questions EitherWilliam E. Janssen, John W. Hiemenz, and Gerald J. ElfenbeinWilliam E. JanssenSearch for more papers by this author, John W. HiemenzSearch for more papers by this author, and Gerald J. ElfenbeinSearch for more papers by this authorPublished Online:8 May 2009https://doi.org/10.1089/scd.1.1993.2.3AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail FiguresReferencesRelatedDetails Volume 2Issue 1Jan 1993 To cite this article:William E. Janssen, John W. Hiemenz, and Gerald J. Elfenbein.When, How, What, and Where to Purge Are Not the Questions Either.Journal of Hematotherapy.Jan 1993.3-5.http://doi.org/10.1089/scd.1.1993.2.3Published in Volume: 2 Issue 1: May 8, 2009PDF download

The SteriCell cell processing instrument is a good choice for a stem cell processing laboratory that is of sufficient size that they cannot share an apheresis machine with the blood bank. It is a laboratory instrument, with no facility for patient connection. Because of its minimal size and weight, it is easily stored in a cramped laboratory. Its automated programs are appropriate for processing of bone marrow and peripheral blood stem cells, and it is quite easy to learn how to use (in our laboratory, most individuals have been completely facile with the SteriCell after fewer than six processings). Based on reported results from other instruments, the SteriCell provides cell yields that are comparable to competing instruments. Service (provided by Haemonetics) has been satisfactory, and support from Terumo has been excellent. We can recommend this instrument to any other laboratory.