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QUANTIFICATION OF CD34+ CELL AND ITS VIABILITY OF FRESH OR CRYOPRESERVED NUCLEATED CELLS BY IMAGE-BASED CELL COUNTER IS COMPARABLE TO STANDARD FLOW CYTOMETER
Author(s):
Young-Ho Lee
,
Young-Ho Lee
Affiliations:
Department of Pediatrics,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of;Blood & Marrow Transplantation Center,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Wee-Jin Rah
,
Wee-Jin Rah
Affiliations:
Department of Pediatrics,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Hani Koh
,
Hani Koh
Affiliations:
Department of Pediatrics,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of;Blood & Marrow Transplantation Center,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Jin Young Suh
,
Jin Young Suh
Affiliations:
Blood & Marrow Transplantation Center,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Hye Jung Eom
,
Hye Jung Eom
Affiliations:
Blood & Marrow Transplantation Center,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Eun-kyung Shin
,
Eun-kyung Shin
Affiliations:
Department of Pediatrics,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Jieun Uhm
,
Jieun Uhm
Affiliations:
Department of Internal Medicine,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of;Blood & Marrow Transplantation Center,HANYANG UNIVERSITY MEDICAL CENTER,Seoul,Korea, Republic Of
Jong Hyun Oh
,
Jong Hyun Oh
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Ji-yeon Lee
,
Ji-yeon Lee
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Sung Rok Bong
,
Sung Rok Bong
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Jee Young Kim
,
Jee Young Kim
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Sunmi Han
,
Sunmi Han
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Chanil Chung
,
Chanil Chung
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Hyun Ju Park
,
Hyun Ju Park
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
Jong Kyu Yoon
Jong Kyu Yoon
Affiliations:
NanoEntek,Seoul,Korea, Republic Of
(Abstract release date: 05/18/17) EHA Library. Lee Y. 05/18/17; 182878; PB2165
Prof. Dr. Young-Ho Lee
Prof. Dr. Young-Ho Lee
Contributions
PDF Abstract
Abstract

Abstract: PB2165

Type: Publication Only

Background
As a standard method for quantification of CD34+ stem cells, flow cytometry has been widely used. However, it has some limitations such as expensive instrumentation, high reagent costs, and poor reproducibility between technicians and laboratories.

Aims

We developed and assessed an instrument performance of a newly-developed image-based microscopic cell counter (ADAM IITM) for enumeration of CD34+ cell and its viability.

Methods
We used samples of fresh and cryopreserved nucleated cells from G-CSF mobilized peripheral blood stem cells (PBSCs) as well as cord blood (CB). We assessed the reproducibility and linearity of the new device and compared numbers and viabilities of CD45+ cells and CD34+ cells determined with the ADAM IITM and flow cytometer.

Results

Each analysis used 10 aliquots from one sample to assess the reproducibility of ADAM IITM, with expected values of 14.77~172.06 CD34+ cells/㎕, 0.08~0.56 CD34(%)/CD45. The number of CD34+ cells determined by ADAM IITM was sufficiently accurate over the expected range, and the intra-assay coefficient of variation (CV) was ≤10.8%. The linearity of CD34+ cell counts was confirmed over a range of dilutions (0.58~280 cells/㎕ of sample). Linearity was satisfactory (R2=0.99). The numbers and viabilities of CD45+ cell and CD34+ cell obtained with the ADAM IITM were highly correlated with those obtained with the flow cytometer (R²>0.9841, p<0.0001). In all samples from fresh/cryopreserved PBSC and fresh/cryopreserved CB, there were no significant differences of total numbers and viabilities of CD45+ cell and CD34+ cell count analyzed from ADAM IITM as well as flow cytometer.

Conclusion

The newly developed image-based microscopic cell counter (ADAM IITM) appears to be suitable for quantification of CD34+ cell and its viability of fresh or cryopreserved PBSCs or CBs.

Session topic: 22. Stem cell transplantation - Clinical

Keyword(s): Peripheral blood stem cell, flow cytometry, cord blood, CD34+ cells

Abstract: PB2165

Type: Publication Only

Background
As a standard method for quantification of CD34+ stem cells, flow cytometry has been widely used. However, it has some limitations such as expensive instrumentation, high reagent costs, and poor reproducibility between technicians and laboratories.

Aims

We developed and assessed an instrument performance of a newly-developed image-based microscopic cell counter (ADAM IITM) for enumeration of CD34+ cell and its viability.

Methods
We used samples of fresh and cryopreserved nucleated cells from G-CSF mobilized peripheral blood stem cells (PBSCs) as well as cord blood (CB). We assessed the reproducibility and linearity of the new device and compared numbers and viabilities of CD45+ cells and CD34+ cells determined with the ADAM IITM and flow cytometer.

Results

Each analysis used 10 aliquots from one sample to assess the reproducibility of ADAM IITM, with expected values of 14.77~172.06 CD34+ cells/㎕, 0.08~0.56 CD34(%)/CD45. The number of CD34+ cells determined by ADAM IITM was sufficiently accurate over the expected range, and the intra-assay coefficient of variation (CV) was ≤10.8%. The linearity of CD34+ cell counts was confirmed over a range of dilutions (0.58~280 cells/㎕ of sample). Linearity was satisfactory (R2=0.99). The numbers and viabilities of CD45+ cell and CD34+ cell obtained with the ADAM IITM were highly correlated with those obtained with the flow cytometer (R²>0.9841, p<0.0001). In all samples from fresh/cryopreserved PBSC and fresh/cryopreserved CB, there were no significant differences of total numbers and viabilities of CD45+ cell and CD34+ cell count analyzed from ADAM IITM as well as flow cytometer.

Conclusion

The newly developed image-based microscopic cell counter (ADAM IITM) appears to be suitable for quantification of CD34+ cell and its viability of fresh or cryopreserved PBSCs or CBs.

Session topic: 22. Stem cell transplantation - Clinical

Keyword(s): Peripheral blood stem cell, flow cytometry, cord blood, CD34+ cells

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