J Korean Med Sci.  1991 Sep;6(3):224-233. 10.3346/jkms.1991.6.3.224.

Requirements for in vitro growth of human thymocytes

• Park SH
• Bae YM
• Park WS
• Kim TJ
• Chung DH
• Song HG
• Cho BJ
• Kim CW
• Lee SK

• KMID: 1421859
• DOI: http://doi.org/10.3346/jkms.1991.6.3.224

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.

Keyword

Human thymocyte; In vitro culure; IL-2receptor expression; Con A receptor

MeSH Terms

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
Antigens, CD
Antigens, CD1
Antigens, CD3
Antigens, Differentiation, T-Lymphocyte
Ionophores
Receptors, Antigen, T-Cell
Tetradecanoylphorbol Acetate