Requirements for in vitro growth of human thymocytes.
10.3346/jkms.1991.6.3.224
- Author:
Seong Hoe PARK
1
;
Young Mee BAE
;
Weon Seo PARK
;
Tae Jin KIM
;
Doo Hyun CHUNG
;
Hyung Geun SONG
;
Byung Jae CHO
;
Chul Woo KIM
;
Sang Kook LEE
Author Information
1. Department of Pathology, College of Medicine, Seoul National University, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Human thymocyte;
In vitro culure;
IL-2receptor expression;
Con A receptor
- MeSH:
Antigens, CD;
Antigens, CD1;
Antigens, CD3;
Antigens, Differentiation, T-Lymphocyte;
Cell Cycle;
Cell Division/drug effects;
Cells, Cultured;
Humans;
Infant;
Infant, Newborn;
Ionophores/pharmacology;
*Lymphocyte Activation/drug effects;
Receptors, Antigen, T-Cell;
T-Lymphocytes/*cytology/drug effects/immunology;
Tetradecanoylphorbol Acetate/pharmacology
- From:Journal of Korean Medical Science
1991;6(3):224-233
- CountryRepublic of Korea
- Language:English
-
Abstract:
Since it is difficult to study human thymocyte maturation in vitro, we have developed an in vitro thymocyte culture system which has allowed us to select the optimal growth conditions for thymocyte subpopulations. Three thymocyte subpopulations (CD3-CD1-, CD1+CD3-, and CD3+CD1-) were isolated by a single step percoll density gradient centrifugation and indirect panning procedure using anti-CD1 and anti-CD3 monoclonal antibodies, and their purity was checked by flow cytometry. The combination of concanavalin A (Con A), tetradecanoylphorbol acetate (TPA), and IL-2 was shown to be the most reliable stimulus for the proliferation of CD3-CD1- thymocytes for up to 15 days in a culture system in vitro. Flow cytometric analysis for the phenotypic change of CD3-CD1- thymocytes revealed a steady increase of CD3 antigen after a 3-day cultivation, whereas there was no change in CD1 antigen intensity. A combination of Con A and IL-2 was both sufficient and necessary to induce growth of CD3+CD1- thymocytes. The major population of immature cortical thymocytes (CD3-CD1+ or CD3+CD1+), which are considered to be the most unresponsive dead-end cells, could not be maintained or stimulated with any combination used in this experiment, even in the presence of thymic accessory cells.
