Ex vivo expansion of umbilical cord blood-derived T-lymphocyte with autologous cord blood plasma

Lee, Sang Soo; Sohn, Woo Seok; Park, Han Moie; Kim, Young Man; Mok, Jung Eun

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.