Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting

Yıl: 2020 Cilt: 3 Sayı: 4 Sayfa Aralığı: 231 - 238 Metin Dili: İngilizce DOI: 10.30714/j-ebr.2020463606 İndeks Tarihi: 14-05-2021

Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting

Öz:
Aim: To compare our parameters as regards: i) cell count via two different automated cell count techniques, and ii) viability via automated trypan blue exclusion and 7-aminoactinomycin D (7-AAD) staining.Method: We used the trypan blue exclusion technique and an automated cell counter and for viability testing, and the trypan blue exclusion technique and the 7-AAD evaluation by flow cytometry. The trypan blue exclusion and the radio frequency techniques were used for automated cell counting. Flow cytometric analysis was performed by evaluating the yielded cellular products for 7-AAD uptake during the cell count of CD34+ cells.Results: The mean values for cell count were estimated as 3.44±1.22x106/ml (range, 2.48-5.71x106/ml) and 4.14±1.94x106/ml (range, 1.77-7.43x106/ml) for the trypan blue exclusion and radio frequency techniques, respectively. Additionally, the mean values for viability analyses via the automated trypan blue exclusion and 7-AAD were 93.38±6.09% (range, 79.00-98.00%) and 99.49±0.60% (range, 98.40-100.00%), respectively.Conclusions: Our study has responded to two fundamental questions: whether the results of both of the automated techniques for cell count correspond with each other, and whether the results of the automated viability assessment conform those of the 7-AAD technique during the manufacturing processes of cellular therapy products intended for clinical use. Even though we have the opportunity to use the hemocytometer in our laboratory setting, the automated trypan blue exclusion technique gives cell count results in concordance within the range of the expectations of our Quality Management System (QMS).
Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
  • [1] Bio-Rad Laboratories, Inc. 2011. TC20™ Automated Cell Counter Instruction Manual Catalog #145-0101.
  • [2] Hsiung F, McCollum T, Hefner E, Rubio T. Comparisons of count reproducibility, accuracy, and time to get results between a hemocytometer and the TC20 automated cell counter. In Bulletin 6003 Rev B; Bio-Rad Laboratories, Inc.: Hercules, CA, 2013;1–4.
  • [3] Kickler TS. Clinical analyzers. Advances in automated cell counting. Anal Chem. 1999;71(12):363R-365R.
  • [4] Sysmex Europe GmbH [Internet]. Sysmex XN-3000. [cited 2019 Dec 24]. Available from: https://www.sysmex.se/products/productsingleview/ xn-3000-967.html.
  • [5] Kwizera R, Akampurira A, Kandole TK, et al. Evaluation of trypan blue stain in the TC20 automated cell counter as a point-ofcare for the enumeration of viable cryptococcal cells in cerebrospinal fluid. Med Mycol. 2018;56(5):559-64.
  • [6] Varan HD, Bay M, Ozturk A, et al. Comparison of the methods evaluating post thawing viability of peripheral blood stem cell graft. Transfus Apher Sci. 2019. 58(2):192-95.
  • [7] Keeney M, Chin-Yee I, Weir K, et al. Single platform flow cytometric absolute CD34+ cell counts based on the ISHAGE guidelines. International Society of Hematotherapy and Graft Engineering. Cytometry. 1998;34(2):61-70.
  • [8] Schmid I, Hausner MA, Cole SW, et al. Simultaneous flow cytometric measurement of viability and lymphocyte subset proliferation. J Immunol Methods. 2001;247(1-2):175-86.
  • [9] Leong CF, Hasbah A, Teh HS, et al. Isolation of purified autologous peripheral blood CD34+ cells with low T cell content using CliniMACS device – a local experience. Malaysian J Pathol. 2008;30(1):31-36.
  • [10] Reich-Slotky R, Colovai AI, Semidei- Pomales M, et al. Determining post-thaw CD34+ cell dose of cryopreserved haematopoietic progenitor cells demonstrates high recovery and confirms their integrity. Vox Sang. 2008;94(4):351- 57.
  • [11] Camacho-Fernández C, Hervás D, Rivas- Sendra A, et al. Comparison of six different methods to calculate cell densities. Plant Methods. 2018;14:30.
  • [12] Kilic P, Bay M, Yildirim Y, et al. A CD34+ cell enrichment protocol of hematopoietic stem cells in a well-established quality management system. Cells Tissues Organs. 2019;207(1):15-20.
  • [13] Miltenyi S, Müller W, Weichel W, et al. High gradient magnetic cell separation with MACS. Cytometry. 1990;11(2):231-38.
  • [14] Arpaci F. CD34 (+) cell selection and purging in hematopoietic cell transplantation. Gülhane Tıp Derg. 2011;53(3): 226-31.
  • [15] Van Dievoet MA, Louagie H, Ghys T. Performance evaluation of the Sysmex(®) XP-300 in an oncology setting: evaluation and comparison of hematological parameters with the Sysmex(®) XN-3000. Int J Lab Hematol. 2016;38(5):490-96.
  • [16] Gratama JW, Kraan J, Keeney M, et al. Validation of the single-platform ISHAGE method for CD34(+) hematopoietic stem and progenitor cell enumeration in an international multicenter study. Cytotherapy. 2003;5(1):55-65.
  • [17] Humpe A, Beck C, Schoch R, et al. Establishment and optimization of a flow cytometric method for evaluation of viability of CD34+ cells after cryopreservation and comparison with trypan blue exclusion staining. Transfusion. 2005;45(7):1208-213.
APA Kilic P, Bay M, OZKAL BAYDIN P, Seker S, Çoşkun Ö, LALEGÜL ÜLKER O, Parmaksiz M, bitirim c, Cevheroğlu O, ÇUBUKÇUOĞLU DENİZ G, ÖZTÜRK A, İPEK Ş, Dalva K, Elcin A, Yilmazer A, Gürman G (2020). Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. , 231 - 238. 10.30714/j-ebr.2020463606
Chicago Kilic Pelin,Bay Meltem,OZKAL BAYDIN PINAR,Seker Sukran,Çoşkun Öznur,LALEGÜL ÜLKER OZGE,Parmaksiz Mahmut,bitirim ceylan verda,Cevheroğlu Orkun,ÇUBUKÇUOĞLU DENİZ GÜNSELİ,ÖZTÜRK AYDIN,İPEK ŞENAY,Dalva Klara,Elcin Ayse Eser,Yilmazer Açelya,Gürman Günhan Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. (2020): 231 - 238. 10.30714/j-ebr.2020463606
MLA Kilic Pelin,Bay Meltem,OZKAL BAYDIN PINAR,Seker Sukran,Çoşkun Öznur,LALEGÜL ÜLKER OZGE,Parmaksiz Mahmut,bitirim ceylan verda,Cevheroğlu Orkun,ÇUBUKÇUOĞLU DENİZ GÜNSELİ,ÖZTÜRK AYDIN,İPEK ŞENAY,Dalva Klara,Elcin Ayse Eser,Yilmazer Açelya,Gürman Günhan Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. , 2020, ss.231 - 238. 10.30714/j-ebr.2020463606
AMA Kilic P,Bay M,OZKAL BAYDIN P,Seker S,Çoşkun Ö,LALEGÜL ÜLKER O,Parmaksiz M,bitirim c,Cevheroğlu O,ÇUBUKÇUOĞLU DENİZ G,ÖZTÜRK A,İPEK Ş,Dalva K,Elcin A,Yilmazer A,Gürman G Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. . 2020; 231 - 238. 10.30714/j-ebr.2020463606
Vancouver Kilic P,Bay M,OZKAL BAYDIN P,Seker S,Çoşkun Ö,LALEGÜL ÜLKER O,Parmaksiz M,bitirim c,Cevheroğlu O,ÇUBUKÇUOĞLU DENİZ G,ÖZTÜRK A,İPEK Ş,Dalva K,Elcin A,Yilmazer A,Gürman G Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. . 2020; 231 - 238. 10.30714/j-ebr.2020463606
IEEE Kilic P,Bay M,OZKAL BAYDIN P,Seker S,Çoşkun Ö,LALEGÜL ÜLKER O,Parmaksiz M,bitirim c,Cevheroğlu O,ÇUBUKÇUOĞLU DENİZ G,ÖZTÜRK A,İPEK Ş,Dalva K,Elcin A,Yilmazer A,Gürman G "Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting." , ss.231 - 238, 2020. 10.30714/j-ebr.2020463606
ISNAD Kilic, Pelin vd. "Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting". (2020), 231-238. https://doi.org/10.30714/j-ebr.2020463606
APA Kilic P, Bay M, OZKAL BAYDIN P, Seker S, Çoşkun Ö, LALEGÜL ÜLKER O, Parmaksiz M, bitirim c, Cevheroğlu O, ÇUBUKÇUOĞLU DENİZ G, ÖZTÜRK A, İPEK Ş, Dalva K, Elcin A, Yilmazer A, Gürman G (2020). Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. Experimental Biomedical Research, 3(4), 231 - 238. 10.30714/j-ebr.2020463606
Chicago Kilic Pelin,Bay Meltem,OZKAL BAYDIN PINAR,Seker Sukran,Çoşkun Öznur,LALEGÜL ÜLKER OZGE,Parmaksiz Mahmut,bitirim ceylan verda,Cevheroğlu Orkun,ÇUBUKÇUOĞLU DENİZ GÜNSELİ,ÖZTÜRK AYDIN,İPEK ŞENAY,Dalva Klara,Elcin Ayse Eser,Yilmazer Açelya,Gürman Günhan Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. Experimental Biomedical Research 3, no.4 (2020): 231 - 238. 10.30714/j-ebr.2020463606
MLA Kilic Pelin,Bay Meltem,OZKAL BAYDIN PINAR,Seker Sukran,Çoşkun Öznur,LALEGÜL ÜLKER OZGE,Parmaksiz Mahmut,bitirim ceylan verda,Cevheroğlu Orkun,ÇUBUKÇUOĞLU DENİZ GÜNSELİ,ÖZTÜRK AYDIN,İPEK ŞENAY,Dalva Klara,Elcin Ayse Eser,Yilmazer Açelya,Gürman Günhan Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. Experimental Biomedical Research, vol.3, no.4, 2020, ss.231 - 238. 10.30714/j-ebr.2020463606
AMA Kilic P,Bay M,OZKAL BAYDIN P,Seker S,Çoşkun Ö,LALEGÜL ÜLKER O,Parmaksiz M,bitirim c,Cevheroğlu O,ÇUBUKÇUOĞLU DENİZ G,ÖZTÜRK A,İPEK Ş,Dalva K,Elcin A,Yilmazer A,Gürman G Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. Experimental Biomedical Research. 2020; 3(4): 231 - 238. 10.30714/j-ebr.2020463606
Vancouver Kilic P,Bay M,OZKAL BAYDIN P,Seker S,Çoşkun Ö,LALEGÜL ÜLKER O,Parmaksiz M,bitirim c,Cevheroğlu O,ÇUBUKÇUOĞLU DENİZ G,ÖZTÜRK A,İPEK Ş,Dalva K,Elcin A,Yilmazer A,Gürman G Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting. Experimental Biomedical Research. 2020; 3(4): 231 - 238. 10.30714/j-ebr.2020463606
IEEE Kilic P,Bay M,OZKAL BAYDIN P,Seker S,Çoşkun Ö,LALEGÜL ÜLKER O,Parmaksiz M,bitirim c,Cevheroğlu O,ÇUBUKÇUOĞLU DENİZ G,ÖZTÜRK A,İPEK Ş,Dalva K,Elcin A,Yilmazer A,Gürman G "Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting." Experimental Biomedical Research, 3, ss.231 - 238, 2020. 10.30714/j-ebr.2020463606
ISNAD Kilic, Pelin vd. "Utility investigation of automated techniques in hematopoietic progenitor cell count and viability assessment in the Good Manufacturing Practice (GMP) setting". Experimental Biomedical Research 3/4 (2020), 231-238. https://doi.org/10.30714/j-ebr.2020463606