Ann Lab Med 2019; 39(4): 388-395  https://doi.org/10.3343/alm.2019.39.4.388
Enumeration of CD34-positive Stem Cells Using the ADAMII Image-based Fluorescence Cell Counter
Haein Yu, B.S.1,2, Jaeeun Yoo, M.D.3, Jung Sil Hwang, B.S.1, Mikyung Kim, B.S.1, Kyung Hee Bae, B.S.1, Dong Wook Jekarl, M.D., Ph.D.3, Jong Hyun Oh, M.S.4, Ji Yeon Lee, M.S.4, Sunmi Han, Ph.D.4, Chanil Chung, Ph.D.4, Myungshin Kim , M.D., Ph.D.1,2,3,*, and Yonggoo Kim , M.D., Ph.D.1,2,3*
1Laboratory Development and Evaluation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea; 2Department of Biomedicine & Health Sciences, Graduate School, The Catholic University of Korea, Seoul, Korea; 3Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea; 4NanoEntek, Seoul, Korea
Corresponding author: Yonggoo Kim, M.D., Ph.D. https://orcid.org/0000-0003-2808-3795
Department of Laboratory Medicine, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul 06591, Korea
Tel: +82-2-2258-1642, Fax: +82-2-2258-1719
E-mail: yonggoo@catholic.ac.kr
Myungshin Kim, M.D., Ph.D. https://orcid.org/0000-0001-8632-0168
Department of Laboratory Medicine, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul 06591, Korea
Tel: +82-2-2258-1645, Fax: +82-2-2258-1719
E-mail: microkim@catholic.ac.kr
*These authors contributed equally to this work.
Received: August 17, 2018; Revised: October 25, 2018; Accepted: February 4, 2019; Published online: July 1, 2019.
© Korean Society for Laboratory Medicine. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background: It is very important to accurately enumerate CD34-positive (CD34+) cells for successful hematopoietic stem cell transplantation (HSCT). We evaluated the ability of the newly developed image based-immunofluorescence cell counter ADAMII (NanoEntek, Seoul, Korea) to enumerate CD34+ cells, which was improved through simultaneous CD45 analysis.
Methods: We enumerated CD34+ cells with ADAMII using 19 peripheral blood (PB) and 91 leukapheresis samples from HSCT donors. Analytical performance, including precision and linearity, was analyzed, and sample stability during storage was evaluated. Viable CD34+ cell count (vCD34) and viable CD45+ cell count (vCD45) and the percentage of viable CD34+ cells among viable CD45+ cells (CD34/CD45) as measured by ADAMII were compared with the corresponding values from two flow cytometry assays, using regression analysis.
Results: ADAMII demonstrated acceptable precision, as CV values of vCD34 from six samples with different counts were all <10% (range: 3.49–9.51%). CV values of the vCD45 and CD34/45 ranged from 4.03% to 9.67% and from 2.48% to 10.07%, respectively. The linearity of vCD34 showed an excellent R2 value (0.99) when analyzed using the intended count and flow cytometry data. The ADAMII and two flow cytometry-based assays generated very similar data for the PB and leukapheresis samples.
Conclusions: ADAMII demonstrated excellent performance for use as a routine clinical assay in terms of CD34+ cell enumeration from PB and leukapheresis samples. Moreover, it could be used as a point-of-care-test for determining mobilization time and predicting an adequate apheresis stem cell product.
Keywords: CD34, Enumeration, Image-based fluorescence cell counter, Flow cytometry, Performance



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