Novel and rapid enumeration method of peripheral blood stem cells using automated hematology analyzer
Corresponding Author
R. Tanosaki
Department of Blood Transfusion and Cellular Therapy, National Cancer Center Hospital, Tokyo, Japan
Correspondence:
Ryuji Tanosaki, Department of Blood Transfusion and Cellular Therapy, National Cancer Center Hospital, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
Tel.: +81 3 3542 2511, +81 3 3547 5201 (X 7066);
Fax: +81 3 3542 3815;
E-mail: [email protected]
Search for more papers by this authorT. Kumazawa
Department of Blood Transfusion and Cellular Therapy, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorA. Nakano
Department of Pathology and Clinical Laboratories, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. Yamagata
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorN. Takahashi
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. Kurosawa
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. W. Kim
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorT. Yamashita
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. Mori
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorY. Heike
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorT. Fukuda
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorH. Tsuda
Department of Pathology and Clinical Laboratories, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorCorresponding Author
R. Tanosaki
Department of Blood Transfusion and Cellular Therapy, National Cancer Center Hospital, Tokyo, Japan
Correspondence:
Ryuji Tanosaki, Department of Blood Transfusion and Cellular Therapy, National Cancer Center Hospital, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
Tel.: +81 3 3542 2511, +81 3 3547 5201 (X 7066);
Fax: +81 3 3542 3815;
E-mail: [email protected]
Search for more papers by this authorT. Kumazawa
Department of Blood Transfusion and Cellular Therapy, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorA. Nakano
Department of Pathology and Clinical Laboratories, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. Yamagata
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorN. Takahashi
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. Kurosawa
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. W. Kim
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorT. Yamashita
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorS. Mori
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorY. Heike
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorT. Fukuda
Department of Hematology and Hematopoietic Stem Cell Transplantation, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorH. Tsuda
Department of Pathology and Clinical Laboratories, National Cancer Center Hospital, Tokyo, Japan
Search for more papers by this authorSummary
Introduction
The number of infused CD34+ cells is crucial to the success of peripheral blood stem cell transplantation (PBSCT). Here, we present, for the first time, a new method of enumerating hematopoietic progenitor cells (HPCs) for PBSCT.
Method
This novel method is based on hemolysis and chemical staining, followed by flow cytometry-based optical detection, conducted using an automated hematology analyzer (XN series, Sysmex). CD34+ cells and HPCs were compared in 76 granulocyte colony-stimulating factor (G-CSF)-mobilized blood or apheresis samples taken from healthy donors (n = 18) or patients undergoing autologous PBSCT (n = 6).
Results
There was a strong correlation between the numbers of HPCs and CD34+ cells (R2 = 0.958). The expected total number of HPCs in the final products, which was estimated from HPCs in pre-apheresis PB or mid-apheresis products, also correlated well with the total number of CD34+ cells in the final products. The change in HPCs in PB closely resembled that of CD34+ cells during mobilization. Experiments using immunomagnetic beads suggested that the majority of CD34+ cells existed in HPCs, and vice versa.
Conclusion
Hematopoietic progenitor cells may serve as surrogates for CD34+ cells in PBSCT. However, further investigations are required to verify this.
References
- 1Civin IC, Strauss LC, Brovall C, Fackler MJ, Schwartz JF, Shaper JH. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-la cells. J Immunol 1984; 133: 157–65.
- 2Bender JG, To LB, Williams S, Schwartzberg LS. Defining a therapeutic dose of peripheral blood stem cells. J Hematother 1992; 1: 329–41.
- 3Bensinger WI, Longin K, Appelbaum F, Rowley S, Weaver C, Lilleby K, Gooley T, Lynch M, Higano T, Klarnet J, Chauncey T, Storb R, Buckner CD. Peripheral blood stem cells (PBSCs) collected after recombinant granulocyte colony stimulating factor (rhG-CSF): an analysis of factors correlating with the tempo of engraftment after transplantation. Br J Haematol 1994; 87: 825–31.
- 4Van den Bossche J, Devreese K, Malfait R, Van de Vyvere M, Wauters A, Neeis H, De Schouwer P. Reference intervals for a complete blood count determined on different automated haematology analysers: Abx Pentra 120 Retic, Coulter Gen-S, Sysmex SE 9500, Abbott Cell Dyn 4000 and Bayer Advia 120. Clin Chem Lab Med 2002; 40: 69–73.
- 5Bourner G, Dhaliwal J, Sumner J. Performance evaluation of the latest fully automated hematology analyzers in a large, commercial laboratory setting: a 4-way, side-by-side study. Lab Hematol 2005; 11: 285–97.
- 6Ishii T, Kawasumi I, Matsumoto H. SE-9000 IMI Channel: focusing on the roles and function of surfactant. Sysmex J International 1997; 7: 123–8.
- 7Takekawa K, Yamane T, Tatsumi N. Determination of hematopoietic stem cells using automated blood cell counter and flow cytometry. Jpn J Clin Pathol 1995; 99: 117–22.
- 8Takekawa K, Yamane T, Hino M, Tatsumi N. Determination of hematopoietic stem cells in peripheral blood by an automated hematology analyzer (SE-9000). Acta Haematol 1998; 100: 130–6.
- 9Endoh A, Yagihashi A, Asanuma K, Moriai R, Izawa A, Koyanagi Y, Sato T, Kobayashi D, Watanabe N. Hematopoietic progenitor cell counts performed by the Sysmex SE-9000 analyzer can guide timing of peripheral blood stem cell harvest. Anticancer Res 2001; 21: 601–4.
- 10Kim MK, Kim S, Jang G, Lee SS, Sym SJ, Lee DH, Kim SW, Jang S, Park CJ, Chi HS, Huh J, Suh C. A randomized comparison of peripheral blood hematopoietic progenitor cell level of 5/mm3 versus 50/mm3 as a surrogate marker to initiate efficient autologous blood stem cell collection. J Clin Apheresis 2007; 22: 277–82.
- 11Lefrere F, Zohar S, Beaudier S, Audat F, Ribeil JA, Ghez D, Varet B, Cavazzana-Calvo M, Dal CL, Letestu R, McIntyre E, Brouzes C. Evaluation of an algorithm based on peripheral blood hematopoietic progenitor cell and CD34+ cell concentrations to optimize peripheral blood progenitor cell collection by apheresis. Transfusion 2007; 47: 1851–7.
- 12Letestu R, Marzac C, Audat F, Belhocine R, Tondeur S, Baccini V, Garçon L, Cortivo LD, Perrot JY, Lefrère F, Valensi F, Ajchenbaum-Cymbalista F. Use of hematopoietic progenitor cell count on the Sysmex XE-2100 for peripheral blood stem cell harvest monitoring. Leuk Lymphoma 2007; 48: 89–96.
- 13Mougi H, Shinmyozu K, Osame M. Determining the optimal time for peripheral blood stem cell harvest by detecting immature cells (immature leukocytes and immature reticulocytes) using two newly developed automatic cell analyzers. Int J Hematol 1997; 66: 303–13.
- 14Oelschlaegel U, Bornhaeuser M, Thiede C, Ehninger G, Hoelig K. HPC enumeration with the Sysmex XE-2100 can guide further flow cytometric CD34(+) measurements and timing of leukaphereses. Cytotherapy 2003; 5: 414–19.
- 15Park KU, Kim SH, Shu C, Kim S, Lee SJ, Park JS, Cho HJ, Kim KW, Lee K, Kim HJ, Park J, Joo MY, Kim JG, Kim T, Lee JH, Kim SB, Kim SW, Lee KH, Lee JS, Kim WK, Park CJ, Chi HS. Correlation of hematopoietic progenitor cell count determined by the SE-automated hematology analyzer with CD34(+) cell count by flow cytometry in leukapheresis products. Am J Hematol 2001; 67: 42–7.
- 16Saigo K, Sugimoto T, Takeuchi S, Kosaka Y, Ryo R, Kumagai S. Estimation of stem cell fractions in peripheral blood stem cell harvest by using an SE-9000 hematology analyzer. Acta Haematol 2000; 103: 157–61.
- 17Lee JL, Kim SB, Lee GW, Ryu MH, Kim EK, Kim S, Kim WK, Lee JS, Park KU, Suh C. Clinical usefulness of the hematopoietic progenitor cell counts in predicting the optimal timing of peripheral blood stem cell harvest. J Korean Med Sci 2003; 18: 27–35.
- 18Peng L, Yang J, Yang H, Peng Z, Xu C, Liu T. Determination of peripheral blood stem cells by the Sysmex SE-9500. Clin Lab Haematol 2001; 23: 231–6.
- 19Suh C, Kim S, Kim SH, Kim EK, Lee JL, Park KU, Lee J, Kim MW, Chi HS, Park CJ, Kim SW. Initiation of peripheral blood progenitor cell harvest based on peripheral blood hematopoietic progenitor cell counts enumerated by the Sysmex SE9000. Transfusion 2004; 44: 1762–8.
- 20Yu J, Leisenring W, Fritschle W, Heimfeld S, Shulman H, Bensinger WI, Holmberg LA, Rowley SD. Enumeration of HPC in mobilized peripheral blood with the Sysmex SE9500 predicts final CD34+ yields in the apheresis collection. Bone Marrow Transplant 2000; 25: 1157–64.
- 21Pollard Y, Watts MJ, Grant D, Chavda N, Linch DC, Machin SJ. Use of the haemopoietic progenitor cell count of the Sysmex SE-9500 to refine apheresis timing of peripheral blood stem cells. Br J Haematol 1999; 106: 538–44.
- 22Vogel W, Kopp HG, Kanz L, Einsele H. Correlations between hematopoietic progenitor cell counts as measured by Sysmex and CD34+ cell harvest yields following mobilization with different regimens. J Cancer Res Clin Oncol 2002; 128: 380–4.
- 23Galembeck E, Alonso A, Meirelles NC. Effects of polyoxyethylene chain length on erythrocyte hemolysis induced by poly[oxyethylene (n) nonylphenol] non-ionic surfactants. Chem Biol Interact 1998; 113: 91–103.
- 24Shalel S, Streichman S, Marmur A. The mechanism of hemolysis by surfactants: effect of solution composition. J Colloid Interface Sci 2002; 252: 66–76.
- 25Sanchez L, Martinez V, Infante MR, Mitjans M, Vinardell MP. Hemolysis and antihemolysis induced by amino acid-based surfactants. Toxicol Lett 2007; 169: 177–84.
- 26Gottfried EL. Lipids of human leukocytes: relation to cell type. J Lipid Res 1967; 8: 321–7.
- 27Gottfried EL. Lipid patterns of leukocytes in health and disease. Semin Hematol 1972; 9: 241–50.
- 28Klock JC, Pieprzyk JK. Cholesterol, phospholipids, and fatty acids of normal immature neutrophils: comparison with acute myeloblastic leukemia cells and normal neutrophils. J Lipid Res 1979; 20: 908–11.
- 29Sutherland DR, Anderson L, Keeney M, Nayar R, Chin-Yee I. The ISHAGE Guidelines for CD34+ cells determination by flow cytometry. J Hematother 1996; 5: 213–26.
- 30Barnett D, Janossy G, Lubenko A, Matutes E, Newland A, Reilly JT. Guideline for the flow cytometric enumeration of CD34+ haematopoietic stem cells. Clin Lab Haematol 1999; 21: 301–8.
- 31Richman CM, Weiner RS, Yankee RA. Increase in circulating stem cells following chemotherapy in man. Blood 1976; 47: 1031–9.
- 32Siena S, Bregni M, Brando B, Ravagnani F, Bonadonna G, Gianni AM. Circulation of CD34+ hematopoietic stem cells in the peripheral blood of high-dose cyclophosphamide-treated patients: enhancement by intravenous recombinant human granulocyte-macrophage colony-stimulating factor. Blood 1989; 74: 1905–14.
- 33Haas R, Mohle R, Fruhauf S, Goldschmidt H, Witt B, Flentje M, Wannenmacher M, Hunstein W. Patient characteristics associated with successful mobilizing and autografting of peripheral blood progenitor cells in malignant lymphoma. Blood 1994; 83: 3787–94.
- 34Fruehauf S, Haas R, Conradt C, Murea S, Witt B, Mohle R, Hunstein W. Peripheral blood progenitor cell (PBPC) counts during steady-state hematopoiesis allow to estimate the yield of mobilized PBPC after Filgrastim (R-metHuG-CSF)-supported cytotoxic chemotherapy. Blood 1995; 85: 2619–26.
- 35Demirer T, Buckner CD, Bensinger WI. Optimization of peripheral blood stem cell mobilization. Stem Cells 1996; 14: 106–16.
- 36Elliott C, Samson DM, Armitage S, Lyttelton MP, McGuigan D, Hargreaves R, Giles C, Abrahamson G, Abboudi Z, Brennan M, Kanfer EJ. When to harvest peripheral-blood stem cells after mobilization therapy: prediction of CD34-positive cell yield by preceding day CD34-positive concentration in peripheral blood. J Clin Oncol 1996; 14: 970–3.
- 37Nakauchi H. Hematopoietic stem cells: are they CD34-positive or CD34-negative? Nat Med 1998; 4: 1009–10.
- 38Bhatia M, Bonnet D, Murdoch B, Gan OI, Dick JE. A newly discovered class of human hematopoietic cells with SCID-repopulating activity. Nat Med 1998; 4: 1038–45.
- 39Goodell MA. CD34+ or CD34−: does it really matter? Blood 1999; 94: 2545–7.
Citing Literature
