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

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

Affiliations
  • 1Laboratory Development and Evaluation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea. yonggoo@catholic.ac.kr, microkim@catholic.ac.kr
  • 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.

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 R 2 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.

Keyword

CD34; Enumeration; Image-based fluorescence cell counter; Flow cytometry; Performance

MeSH Terms

Blood Component Removal
Cell Count*
Flow Cytometry
Fluorescence*
Hematopoietic Stem Cell Transplantation
Humans
Leukapheresis
Seoul
Stem Cells*
Tissue Donors

Figure

  • Fig. 1 ADAMII-CD34 assay procedure.

  • Fig. 2 Linearity of viable CD34-positive cell count (vCD34) using two samples. vCD34 counts compared with intended values (A, C) and FACSCanto II values (B, D).

  • Fig. 3 Stability results of ADAMII for viable CD45-positive cell count (vCD45) (A) and viable CD34-positive cell count (vCD34) (B), along with box plots showing percentage of recovery (C, D).

  • Fig. 4 Correlations between vCD45, vCD34, and CD34/CD45 in ADAMII and flow cytometry, shown through Passing–Bablok regression analyses (A–F) and Bland–Altman plots (G–L). Data from flow cytometry including FACSCanto II and FACSAria II on the horizontal (x) axis. Data from ADAMII and their differences from flow cytometry on the vertical (y) axis.Abbreviations: vCD45, viable CD45-positive cell count; vCD34, viable CD34-positive cell count; CD34/CD45, percentage of viable CD34-positive cells among viable CD45-positive cells.


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