Ex vivo expansion of umbilical cord blood-derived T-lymphocyte with autologous cord blood plasma.
- Author:
Sang Soo LEE
1
;
Woo Seok SOHN
;
Han Moie PARK
;
Young Man KIM
;
Jung Eun MOK
Author Information
1. Department of Obstetrics and Gynecology, University of Ulsan, Gangneung Asan Hospital, Gangneung, Korea.
- Publication Type:Original Article
- Keywords:
Umbilical cord blood;
T-lymphocyte;
Ex vivo expansion;
Autologous cord blood plasma;
Adoptive cellular immunotherapy
- MeSH:
Centrifugation;
Fetal Blood*;
Immunotherapy, Adoptive;
Interleukin-2;
Plasma*;
T-Lymphocytes*;
Umbilical Cord*
- From:Korean Journal of Obstetrics and Gynecology
2005;48(3):607-616
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
OBJECTIVE: The objective of this study was to establish a clinically applicable culture system by investigating the use of autologous cord blood plasma (ACBP) instead of fetal bovine serum (FBS) for the ex vivo expansion of umbilical cord blood (UCB) T-lymphocytes. METHODS: Fresh UCB mononuclear cell (MNC) fractions were isolated by Ficoll-Hypaque density centrifugation. The nonadherent MNC fractions were then cultured with the anti-CD3 antibody 5 microgram/mL plus IL-2 175 U/mL in the presence of 10% FBS, 10% ACBP or homologous cord blood plasma (HCBP). On day 8, proliferation rate, cell surface markers, cytotoxic assay of UCB T-lymphocytes according to the medium supplemented with FBS, ACBP or HCBP were evaluated. RESULTS: Proliferation studies demonstrated a significant increase in the proliferative ability of UCB T-lymphocytes incubated in anti- CD3 and IL-2 irrespective of the medium supplemented with FBS or ACBP. In the FBS supplemented medium, expressions of the activated T-lymphocytes were increased significantly after culture: CD3+CD8+, CD3+CD25+, CD3+CD38+, and CD45RO+ (p<0.05). Also in the ACBP supplemented medium, expressions of the activated T-lymphocytes were increased significantly after culture: CD3+ CD8+, CD3+CD25+, and CD45RO+ (p<0.05). In the HCBP supplemented medium, expressions of the activated T-lymphocytes were increased significantly after culture as in the ACBP: CD3+CD8+, CD3+CD25+, and CD45RO+ (p<0.05). Of the activated T-lymphocytes, increase of cytotoxic CD3+CD8+ cells increased significantly in the ACBP and HCBP groups compared to FBS group (p<0.05). CONCLUSION: These findings support the feasibility of ex vivo expansion of umbilical cord blood T-lymphocytes in the medium supplemented with autologous cord blood plasma, instead of fetal bovine serum, for future adoptive cellular immunotherapy.
