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Ann Lab Med. 2019 Jul;39(4):388-395. doi: 10.3343/alm.2019.39.4.388.

Enumeration of CD34-positive Stem Cells Using the ADAMII Image-based Fluorescence Cell Counter.

Yu H1,2, Yoo J3, Hwang JS1, Kim M1, Bae KH1, Jekarl DW3, Oh JH4, Lee JY4, Han S4, Chung C4, Kim M#1,2,5, Kim Y#1,2,6.

Author information

1
Laboratory Development and Evaluation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea.
2
Department of Biomedicine & Health Sciences, Graduate School, The Catholic University of Korea, Seoul, Korea.
3
Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea.
4
NanoEntek, Seoul, Korea.
5
Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea. microkim@catholic.ac.kr.
6
Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea. yonggoo@catholic.ac.kr.
#
Contributed equally

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; Flow cytometry; Image-based fluorescence cell counter; Performance

PMID:
30809985
PMCID:
PMC6400716
[Available on 2019-07-01]
DOI:
10.3343/alm.2019.39.4.388
[Indexed for MEDLINE]
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