Korean J Hematol 2004; 39(4):

Published online December 31, 2004

© The Korean Society of Hematology

골수, 말초혈액 및 제대혈 단일 CD34+ 세포의 성장 및 분화능 비교 연구

신명근, 조덕, 이진솔, 최현우, 오봉준, 기승정, 신종희, 서순팔, 양동욱, 김수현

전남대학교 의과대학 진단검사의학교실
, 서남대학교 의과대학 진단검사의학교실

Plating Efficiency of Single CD34+ Cell from Bone Marrow, Peripheral Blood and Cord Blood.

Myung Geun Shin, Duck Cho, Jin Sol Lee, Hyun Woo Choi, Bong Joon Oh, Soo Hyun Kim, Seung Jung Kee, Jong Hee Shin, Soon Pal Suh, Dong Wook Ryang

Department of Laboratory Medicine, Chonnam National University Medical School, Gwangju, Korea,
College of Medicine, Seonam University, Namwon, Korea.

Abstract

Background :
Although bone marrow (BM) CD34+cells and peripheral blood (PB) CD34+ cells are developmentally and functionally related, the recent data suggested that they may have different functional and clinical capabilities. Moreover, they have differential gene expression underlying the functional distinctions of primary human CD34+ hematopoietic stem and progenitor cells from BM and PB. The aim of this study was to investigate the plating efficiency of single CD34+ progenitor cell from BM, PB and umbilical cord blood (UCB).
Methods :
After sorting, single CD34+ cells were cultured in individual wells of 96-well plates in serum-free medium containing selected hematopoietic growth factors, with or without G-CSF. Plating efficiency was microscopically determined by the presence of clusters of viable cells:[the number of positive (cells were present) wells/total wells]x100. CD34+ cell-derived colonies were classified according to the cell number per well.
Result :
Although there was some variation of plating efficiency of CD34+ cells among six normal BMs, six PBs and five UCBs, overall average plating efficiency of single CD34+ cells from BM, PB and UCB was 30% (30.0+/-11.7, mean+/-SD), 79% (78.6+/-11.7) and 45% (45.3+/-9.3) respectively. As expected, the colony size was increased in the presence of G-CSF.
Conclusion :
The results of this study clearly showed that the different ex vivo expansion of single CD34+ progenitor cells from BM, PB and UCB. These might be an important data for understanding stem cell expansion in vivo and designing clinical application.

Keywords Single CD34+ cell, Plating efficiency, Bone marrow, Peripheral Blood, Umbilical cord blood

Article

Korean J Hematol 2004; 39(4): 217-222

Published online December 31, 2004

Copyright © The Korean Society of Hematology.

골수, 말초혈액 및 제대혈 단일 CD34+ 세포의 성장 및 분화능 비교 연구

신명근, 조덕, 이진솔, 최현우, 오봉준, 기승정, 신종희, 서순팔, 양동욱, 김수현

전남대학교 의과대학 진단검사의학교실
, 서남대학교 의과대학 진단검사의학교실

Plating Efficiency of Single CD34+ Cell from Bone Marrow, Peripheral Blood and Cord Blood.

Myung Geun Shin, Duck Cho, Jin Sol Lee, Hyun Woo Choi, Bong Joon Oh, Soo Hyun Kim, Seung Jung Kee, Jong Hee Shin, Soon Pal Suh, Dong Wook Ryang

Department of Laboratory Medicine, Chonnam National University Medical School, Gwangju, Korea,
College of Medicine, Seonam University, Namwon, Korea.

Abstract

Background :
Although bone marrow (BM) CD34+cells and peripheral blood (PB) CD34+ cells are developmentally and functionally related, the recent data suggested that they may have different functional and clinical capabilities. Moreover, they have differential gene expression underlying the functional distinctions of primary human CD34+ hematopoietic stem and progenitor cells from BM and PB. The aim of this study was to investigate the plating efficiency of single CD34+ progenitor cell from BM, PB and umbilical cord blood (UCB).
Methods :
After sorting, single CD34+ cells were cultured in individual wells of 96-well plates in serum-free medium containing selected hematopoietic growth factors, with or without G-CSF. Plating efficiency was microscopically determined by the presence of clusters of viable cells:[the number of positive (cells were present) wells/total wells]x100. CD34+ cell-derived colonies were classified according to the cell number per well.
Result :
Although there was some variation of plating efficiency of CD34+ cells among six normal BMs, six PBs and five UCBs, overall average plating efficiency of single CD34+ cells from BM, PB and UCB was 30% (30.0+/-11.7, mean+/-SD), 79% (78.6+/-11.7) and 45% (45.3+/-9.3) respectively. As expected, the colony size was increased in the presence of G-CSF.
Conclusion :
The results of this study clearly showed that the different ex vivo expansion of single CD34+ progenitor cells from BM, PB and UCB. These might be an important data for understanding stem cell expansion in vivo and designing clinical application.

Keywords: Single CD34+ cell, Plating efficiency, Bone marrow, Peripheral Blood, Umbilical cord blood

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