CD34+ Hematopoietik Kök Hücrelerin Cryofit® DMSO Kullanılarak Kriyoprezervasyonu ve Sonuçları
Year 2025,
Volume: 7 Issue: 2, 273 - 279, 23.06.2025
Semih Başcı
,
Hikmettullah Batgi
,
Sinem Namdaroğlu
Abstract
Amaç: Otolog hematopoetik kök hücre nakli (OKHN), hematolojik maligniteler ve immün bozukluklar için önemli bir tedavi yöntemidir. CD34+ hematopoetik kök hücrelerin (HKH) kriyoprezervasyonu, nakil başarısını sağlamak için kritik bir adımdır. Dimetil sülfoksit (DMSO), yaygın olarak kullanılan bir kriyoprotektandır ancak infüzyona bağlı toksisiteler oluşturabilir. CryoFit® DMSO, hücre canlılığını artırmayı ve olumsuz etkileri azaltmayı amaçlamaktadır. Bu çalışma, DMSO ile kriyoprezervasyonun OKHN’de etkinliğini ve güvenliğini değerlendirmektedir.
Gereç ve Yöntem: Bu çalışma, tek merkezli retrospektif bir analiz olup daha önceden CryoFit® DMSO ile kriyoprezervasyon yapılmış kök hücrelerden OKHN olan 80 hasta dahil edildi. Kök hücre mobilizasyonu, granülosit koloni stimüle edici faktör (G-CSF) ± kemoterapi ve gerekli durumlarda plerixafor kullanılarak gerçekleştirilmiştir. CD34+ hücreleri kriyoprezervasyondan önce akış sitometrisi ile sayıldı. Nakil sonrası kök hücre yamanması (engraftment), transfüzyon ihtiyacı ve infüzyona bağlı yan etkiler değerlendirildi. Veri analizi SPSS 26.0 yazılımı ile yapıldı.
Bulgular: Hastaların medyan yaşı 58.5 (aralık: 19-75), ve kohortun %53,8’i kadındı. En yaygın tanı multipl miyelomdu (%57,5). Toplanan CD34+ hücrelerin medyan miktarı 5.8 x 106/kg (aralık: 3.2-14) idi. Çözündürme sonrası hücre canlılığı %98 (aralık: %90-99.5) olarak bulundu. Nötrofil ve trombosit engraftmanı sırasıyla medyan 13 ve 17 gün olarak izlendi. Medyan hastanede kalış süresi 24 gün (aralık: 15-60) idi. Hastaların %26,3’ünde infüzyona bağlı yan etkiler gözlendi, en yaygın olanı bulantı/kusmaydı (%15) ve tüm yan etkiler yönetilebilir düzeydeydi.
Sonuç: CryoFit® DMSO, CD34+ HKH’ların çözündürme sonrası yüksek canlılık oranları ile etkin bir şekilde korunmasını sağlamıştır. Hafif infüzyona bağlı toksisiteler gözlemlenmiş ancak geçici olmuştur. Bu sonuçlar, CryoFit® DMSO’nun OKHN’de kullanımını desteklemektedir. Kriyoprezervasyon protokollerini optimize etmek için çok merkezli ileri çalışmalar gereklidir.
References
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Copelan EA. Hematopoietic stem-cell transplantation. N Engl J Med 2006;354:1813-1826.
-
Logue M, Savani BN. Understanding basic steps to hematopoietic stem cell transplantation evaluation. Am J Blood Res 2013;3:102-106.
-
Berebichez-Fridman R, Montero-Olvera PR. Sources and clinical applications of mesenchymal stem cells: state-of-the-art review. Sultan Qaboos Univ Med J 2018;18(3):e264-e277.
-
Cottler-Fox MH, Lapidot T, Petit I et al. Stem cell mobilization. Hematology Am Soc Hematol Educ Program 2003:419-437.
-
Mitrus I, Smagur A, Giebel S et al. A faster reconstitution of hematopoiesis after autologous transplantation of hematopoietic cells cryopreserved in 7.5% dimethyl sulfoxide if compared to 10% dimethyl sulfoxide containing medium. Cryobiology 2013;67:327-331.
-
Allan DS, Keeney M, Popma J et al. Peripheral blood stem cells: number of viable CD34+ cells reinfused predicts engraftment in autologous hematopoietic stem cell transplantation summary. Bone Marrow Transplant 2002;29:967-972.
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World Health Organization. Guidance on Regulations for the Transport of Infectious Substances 2013-2014. World Health Organization 2013:6-7.
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Jansen J, Nolan PL, Reeves MI et al. Transportation of peripheral blood progenitor cell products: effects of time, temperature and cell concentration. Cytotherapy 2009;11:79-85.
-
Kao GS, Kim HT, Daley H, et al. Validation of short-term handling and storage conditions for marrow and peripheral blood stem cell products. Transfusion 2011;51:137-145.
-
Kardduss-Urueta A, Gale RP, Gutierrez-Aguirre CH et al. Freezing the graft is not necessary for autotransplants for plasma cell myeloma and lymphomas. Bone Marrow Transplant 2018;53:457-460.
-
Moreb J, Byrne M, Salmasinia D, et al. Tandem autologous stem cell transplantation for multiple myeloma patients based on response to their first transplant-a prospective phase II study. Clin Med Insights Oncol 2014;8:101-105.
-
Vrhovac R, Perić Z, Jurenec S et al. Post-thaw viability of cryopreserved hematopoietic progenitor cell grafts: does it matter? Coll Antropol 2010;34(1):163-169.
-
Felix OMWO, Tunes G, Ginani VC, et al. The influence of cell concentration at cryopreservation on neutrophil engraftment after autologous peripheral blood stem cell transplantation. Hematol Transfus Cell Ther 2018;40:233-239.
-
Gonçalves TL, Benvegnú DM, Bonfanti G. Specific factors influence the success of autologous and allogeneic hematopoietic stem cell transplantation. Oxid Med Cell Longev 2009;2:82-87.
-
Guttridge M, Soh T, Belfield H, Sidders C, Watt S. Storage time affects umbilical cord blood viability. Transfusion 2014;54:1278-1285.
-
Donmez A, Yilmaz F, Soyer N, Cagirgan S, Arik B, Tombuloglu M. The loss of CD34+ cells in peripheral hematopoietic stem cell products cryopreserved by noncontrolled rate freezing and stored at -80°C after overnight storage. Transfus Apher Sci 2014;51:188-192.
-
Watz E, Remberger M, Ringden O et al. Quality of the hematopoietic stem cell graft affects the clinical outcome of allogeneic stem cell transplantation. Transfusion 2015;55:2339-2350.
-
Morgenstern DA, Ahsan G, Brocklesby M et al. Post-thaw viability of cryopreserved peripheral blood stem cells (PBSC) does not guarantee functional activity: important implications for quality assurance of stem cell transplant programs. Br J Haematol 2016;174:942-951.
-
Bittencourt MCB, Mariano L, Moreira F, Schmidt-Filho J, Mendrone-Jr A, Rocha V. Cryopreserved versus non-cryopreserved peripheral blood stem cells for autologous transplantation after high-dose melphalan in multiple myeloma: comparative analysis. Bone Marrow Transplant 2019;54:138-141.
-
Donmez A, Tombuloglu M, Gungor A, Soyer N, Saydam G, Cagirgan S. Clinical side effects during peripheral blood progenitor cell infusion. Transfus Apher Sci 2007;36:95-101.
-
Shu Z, Heimfeld S, Gao D. Hematopoietic SCT with cryopreserved grafts: adverse reactions after transplantation and cryoprotectant removal before infusion. Bone Marrow Transplant 2014;49:469-476.
-
Santos NCF-CJ. Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular and molecular aspects. Biochem Pharmacol 2003;65:1035-1041.
-
Hoyt R, Szer J, Grigg A. Neurological events associated with the infusion of cryopreserved bone marrow and/or peripheral blood progenitor cells. Bone Marrow Transplant 2000;25:1285-1287.
-
Alessandrino EP, Bernasconi P, Caldera D et al. Adverse events occurring during bone marrow or peripheral blood progenitor cell infusion: analysis of 126 cases. Bone Marrow Transplant 1999;23:533-537.
-
Sutherland DR, Anderson L, Keeney M, Nayar R, Chin-Yee I. The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering. J Hematother 1996;5(3):213-226.
-
Awan M, Buriak I, Fleck R et al. Dimethyl sulfoxide: a central player since the dawn of cryobiology, is efficacy balanced by toxicity? Regen Med 2020;15(3):1463-1491.
-
Giralt S, Costa L, Schriber J et al. Optimizing autologous stem cell mobilization strategies to improve patient outcomes: consensus guidelines and recommendations. Biol Blood Marrow Transplant 2014;20(3):295-308.
-
Otrock ZK, Sempek DS, Carey S, Grossman BJ. Adverse events of cryopreserved hematopoietic stem cell infusions in adults: a single-center observational study. Transfusion 2017;57(6):1522-1526.
-
Mitrus I, Smagur A, Fidyk W et al. Reduction of DMSO concentration in cryopreservation mixture from 10% to 7.5% and 5% has no impact on engraftment after autologous peripheral blood stem cell transplantation: results of a prospective, randomized study. Bone Marrow Transplant 2018;53(3):274-280.
Cryopreservation of CD34+ Hematopoietic Stem Cells Using Cryofit® DMSO and Its Outcomes
Year 2025,
Volume: 7 Issue: 2, 273 - 279, 23.06.2025
Semih Başcı
,
Hikmettullah Batgi
,
Sinem Namdaroğlu
Abstract
Objective: Autologous hematopoietic stem cell transplantation (auto-SCT) is a key treatment for hematological malignancies and immune disorders. Cryopreservation of CD34+ hematopoietic stem cells (HSCs) ensures transplant success. Dimethyl sulfoxide (DMSO) is a widely used cryoprotectant but can cause infusion-related toxicities. CryoFit® DMSO aims to enhance cell viability while reducing adverse effects. This study evaluates its efficacy and safety in auto-SCT.
Material and Method: A single-center, retrospective study was conducted on 80 patients who underwent auto- SCT with CD34+ HSCs cryopreserved using CryoFit® DMSO. Mobilization was performed using granulocyte colony-stimulating factor (G-CSF) ± chemotherapy and plerixafor when required. CD34+ cells were quantified via flow cytometry before cryopreservation. Post-transplant engraftment, transfusion needs, and infusion-related side effects were assessed. Data analysis was conducted using SPSS 26.0.
Results: The median patient age was 58.5 years (range: 19-75) and 53.8% (n=43) of the cohort sample was female. Multiple myeloma was the most common diagnosis (57.5%). The median collected CD34+ cell count was 5.8 x 106/ kg (range: 3.2-14). Post-thaw viability was 98% (range: 90-99.5%). Neutrophil and platelet engraftment occurred at medians of 13 and 17 days, respectively. The median hospitalization duration was 24 days (range: 15-60). Infusion-related adverse effects occurred in 26.3% of patients, primarily nausea/vomiting (15%), all manageable.
Conclusion: CryoFit® DMSO effectively preserves CD34+ HSCs with high post-thaw viability and favorable engraftment. Mild infusion-related toxicities were observed but were transient. The results support its continued use in auto-SCT. Further multicenter studies are required to optimize cryopreservation protocols.
Ethical Statement
Approval for the study was obtained from the Dokuz Eylül University Non-Interventional Research Ethics Committee on 06/12/2023. Decision no: 2023/39-12.
References
-
Copelan EA. Hematopoietic stem-cell transplantation. N Engl J Med 2006;354:1813-1826.
-
Logue M, Savani BN. Understanding basic steps to hematopoietic stem cell transplantation evaluation. Am J Blood Res 2013;3:102-106.
-
Berebichez-Fridman R, Montero-Olvera PR. Sources and clinical applications of mesenchymal stem cells: state-of-the-art review. Sultan Qaboos Univ Med J 2018;18(3):e264-e277.
-
Cottler-Fox MH, Lapidot T, Petit I et al. Stem cell mobilization. Hematology Am Soc Hematol Educ Program 2003:419-437.
-
Mitrus I, Smagur A, Giebel S et al. A faster reconstitution of hematopoiesis after autologous transplantation of hematopoietic cells cryopreserved in 7.5% dimethyl sulfoxide if compared to 10% dimethyl sulfoxide containing medium. Cryobiology 2013;67:327-331.
-
Allan DS, Keeney M, Popma J et al. Peripheral blood stem cells: number of viable CD34+ cells reinfused predicts engraftment in autologous hematopoietic stem cell transplantation summary. Bone Marrow Transplant 2002;29:967-972.
-
World Health Organization. Guidance on Regulations for the Transport of Infectious Substances 2013-2014. World Health Organization 2013:6-7.
-
Jansen J, Nolan PL, Reeves MI et al. Transportation of peripheral blood progenitor cell products: effects of time, temperature and cell concentration. Cytotherapy 2009;11:79-85.
-
Kao GS, Kim HT, Daley H, et al. Validation of short-term handling and storage conditions for marrow and peripheral blood stem cell products. Transfusion 2011;51:137-145.
-
Kardduss-Urueta A, Gale RP, Gutierrez-Aguirre CH et al. Freezing the graft is not necessary for autotransplants for plasma cell myeloma and lymphomas. Bone Marrow Transplant 2018;53:457-460.
-
Moreb J, Byrne M, Salmasinia D, et al. Tandem autologous stem cell transplantation for multiple myeloma patients based on response to their first transplant-a prospective phase II study. Clin Med Insights Oncol 2014;8:101-105.
-
Vrhovac R, Perić Z, Jurenec S et al. Post-thaw viability of cryopreserved hematopoietic progenitor cell grafts: does it matter? Coll Antropol 2010;34(1):163-169.
-
Felix OMWO, Tunes G, Ginani VC, et al. The influence of cell concentration at cryopreservation on neutrophil engraftment after autologous peripheral blood stem cell transplantation. Hematol Transfus Cell Ther 2018;40:233-239.
-
Gonçalves TL, Benvegnú DM, Bonfanti G. Specific factors influence the success of autologous and allogeneic hematopoietic stem cell transplantation. Oxid Med Cell Longev 2009;2:82-87.
-
Guttridge M, Soh T, Belfield H, Sidders C, Watt S. Storage time affects umbilical cord blood viability. Transfusion 2014;54:1278-1285.
-
Donmez A, Yilmaz F, Soyer N, Cagirgan S, Arik B, Tombuloglu M. The loss of CD34+ cells in peripheral hematopoietic stem cell products cryopreserved by noncontrolled rate freezing and stored at -80°C after overnight storage. Transfus Apher Sci 2014;51:188-192.
-
Watz E, Remberger M, Ringden O et al. Quality of the hematopoietic stem cell graft affects the clinical outcome of allogeneic stem cell transplantation. Transfusion 2015;55:2339-2350.
-
Morgenstern DA, Ahsan G, Brocklesby M et al. Post-thaw viability of cryopreserved peripheral blood stem cells (PBSC) does not guarantee functional activity: important implications for quality assurance of stem cell transplant programs. Br J Haematol 2016;174:942-951.
-
Bittencourt MCB, Mariano L, Moreira F, Schmidt-Filho J, Mendrone-Jr A, Rocha V. Cryopreserved versus non-cryopreserved peripheral blood stem cells for autologous transplantation after high-dose melphalan in multiple myeloma: comparative analysis. Bone Marrow Transplant 2019;54:138-141.
-
Donmez A, Tombuloglu M, Gungor A, Soyer N, Saydam G, Cagirgan S. Clinical side effects during peripheral blood progenitor cell infusion. Transfus Apher Sci 2007;36:95-101.
-
Shu Z, Heimfeld S, Gao D. Hematopoietic SCT with cryopreserved grafts: adverse reactions after transplantation and cryoprotectant removal before infusion. Bone Marrow Transplant 2014;49:469-476.
-
Santos NCF-CJ. Multidisciplinary utilization of dimethyl sulfoxide: pharmacological, cellular and molecular aspects. Biochem Pharmacol 2003;65:1035-1041.
-
Hoyt R, Szer J, Grigg A. Neurological events associated with the infusion of cryopreserved bone marrow and/or peripheral blood progenitor cells. Bone Marrow Transplant 2000;25:1285-1287.
-
Alessandrino EP, Bernasconi P, Caldera D et al. Adverse events occurring during bone marrow or peripheral blood progenitor cell infusion: analysis of 126 cases. Bone Marrow Transplant 1999;23:533-537.
-
Sutherland DR, Anderson L, Keeney M, Nayar R, Chin-Yee I. The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering. J Hematother 1996;5(3):213-226.
-
Awan M, Buriak I, Fleck R et al. Dimethyl sulfoxide: a central player since the dawn of cryobiology, is efficacy balanced by toxicity? Regen Med 2020;15(3):1463-1491.
-
Giralt S, Costa L, Schriber J et al. Optimizing autologous stem cell mobilization strategies to improve patient outcomes: consensus guidelines and recommendations. Biol Blood Marrow Transplant 2014;20(3):295-308.
-
Otrock ZK, Sempek DS, Carey S, Grossman BJ. Adverse events of cryopreserved hematopoietic stem cell infusions in adults: a single-center observational study. Transfusion 2017;57(6):1522-1526.
-
Mitrus I, Smagur A, Fidyk W et al. Reduction of DMSO concentration in cryopreservation mixture from 10% to 7.5% and 5% has no impact on engraftment after autologous peripheral blood stem cell transplantation: results of a prospective, randomized study. Bone Marrow Transplant 2018;53(3):274-280.