Korean J Hematol 2004; 39(2):

Published online June 30, 2004

© The Korean Society of Hematology

증폭된 조혈전구세포에서 Fas 항원 및 Bax의 발현과 아포토시스

김찬규, 이남수, 배상범, 이규택, 박성규, 원종호, 박희숙, 홍대식, 장금하, 정희정, 김숙자

순천향대학교 의과대학 내과학교실, 임상분자생물학연구소

The Expression of Fas antigen and Bax and Apoptosis in Ex Vivo Expanded Hematopoietic Progenitor Cells

Chan Kyu Kim, Nam Su Lee, Sang Byung Bae, Kyu Tack Lee, Sung Kyu Park, Jong Ho Won, Hee Sook Park, Dae Sik Hong, Kum Ha Jang, Hee Jeong Cheong, Sook Ja Kim

Department of Internal Medicine and Institute for Clinical Biology Research, Soonchunhyang University College of Medicine, Bucheon, Korea

Abstract

BACKGROUND:
During ex vivo expansion of cord blood (CB) CD34+ cells, differentiation of the expanded cells happened and hematopoietic potential of the progenitor cells decreased. In this study, we evaluate the effect of the expression of Fas antigen, Bcl-2, and Bax on CD34+ or AC133+ hematopoietic progenitor cells during ex vivo expansion.
METHODS : CD34+ and AC133+ cells isolated from human CB were cultured in serum free medium supplemented with several cytokines for 7 days. After expansion culture, we re isolated CD34+ and AC133+ cells and compared the numbers of granulocyte-macrophage colony- forming units (CFU-GM) and granulocyte, erythrocyte, monocyte, and macrophage colony-forming units (CFU-GEMM), and expression of Fas antigen, Bcl-2, and Bax with unexpanded cells.
RESULTS : CFU-GM was expanded 23.94 fold in CD34+ cells and 15.22 fold in AC133+ cells at day 7 of culture but CFU-GEMM was not expanded. The expression of Fas antigen and Bax was 7.44% and 2.75%, respectively, in fresh isolated CD34+ cells and increased to 19.71 % and 33.67%, respectively, in expanded CD34+ cells at day 7 culture, but Bcl-2 was not changed. In case of AC133+ cells, the expression of Fas antigen and Bax were also increased from 5.87% and 6.19% to 24.85% and 22.83%, respectively, and Bcl-2 was slightly decreased. Apoptosis was not changed in CD34+ cells and AC133+ cells during ex vivo expansion.
CONCLUSION :
These results indicate that the nature of expansion was similar between CD34+ and AC133+ cells, and expression of Fas antigen and Bax increased on CD34+ and AC133+ cells during ex vivo expansion. Selection of the expanded progenitor cells without apoptosis may be useful for the hematopoietic stem cell transplantation.

Keywords Apoptosis, Fas antigen, Bax, ex vivo expansion, Hematopoietic progenitor cells

Article

Korean J Hematol 2004; 39(2): 95-102

Published online June 30, 2004

Copyright © The Korean Society of Hematology.

증폭된 조혈전구세포에서 Fas 항원 및 Bax의 발현과 아포토시스

김찬규, 이남수, 배상범, 이규택, 박성규, 원종호, 박희숙, 홍대식, 장금하, 정희정, 김숙자

순천향대학교 의과대학 내과학교실, 임상분자생물학연구소

The Expression of Fas antigen and Bax and Apoptosis in Ex Vivo Expanded Hematopoietic Progenitor Cells

Chan Kyu Kim, Nam Su Lee, Sang Byung Bae, Kyu Tack Lee, Sung Kyu Park, Jong Ho Won, Hee Sook Park, Dae Sik Hong, Kum Ha Jang, Hee Jeong Cheong, Sook Ja Kim

Department of Internal Medicine and Institute for Clinical Biology Research, Soonchunhyang University College of Medicine, Bucheon, Korea

Abstract

BACKGROUND:
During ex vivo expansion of cord blood (CB) CD34+ cells, differentiation of the expanded cells happened and hematopoietic potential of the progenitor cells decreased. In this study, we evaluate the effect of the expression of Fas antigen, Bcl-2, and Bax on CD34+ or AC133+ hematopoietic progenitor cells during ex vivo expansion.
METHODS : CD34+ and AC133+ cells isolated from human CB were cultured in serum free medium supplemented with several cytokines for 7 days. After expansion culture, we re isolated CD34+ and AC133+ cells and compared the numbers of granulocyte-macrophage colony- forming units (CFU-GM) and granulocyte, erythrocyte, monocyte, and macrophage colony-forming units (CFU-GEMM), and expression of Fas antigen, Bcl-2, and Bax with unexpanded cells.
RESULTS : CFU-GM was expanded 23.94 fold in CD34+ cells and 15.22 fold in AC133+ cells at day 7 of culture but CFU-GEMM was not expanded. The expression of Fas antigen and Bax was 7.44% and 2.75%, respectively, in fresh isolated CD34+ cells and increased to 19.71 % and 33.67%, respectively, in expanded CD34+ cells at day 7 culture, but Bcl-2 was not changed. In case of AC133+ cells, the expression of Fas antigen and Bax were also increased from 5.87% and 6.19% to 24.85% and 22.83%, respectively, and Bcl-2 was slightly decreased. Apoptosis was not changed in CD34+ cells and AC133+ cells during ex vivo expansion.
CONCLUSION :
These results indicate that the nature of expansion was similar between CD34+ and AC133+ cells, and expression of Fas antigen and Bax increased on CD34+ and AC133+ cells during ex vivo expansion. Selection of the expanded progenitor cells without apoptosis may be useful for the hematopoietic stem cell transplantation.

Keywords: Apoptosis, Fas antigen, Bax, ex vivo expansion, Hematopoietic progenitor cells

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