Production method for cell populations
The present invention addresses the problem of providing cell populations having a high proportion of CD4-positive naive T cells and/or CD4-positive central memory T cells, and a production method thereof. The present invention provides a production method for CD4-positive T cell populations which is characterized by using anti-CD3 antibodies, fibronectin fragments, and Interleukin-4. The method is characterized not only by the attainment of a cell group with a high proportion of CD4-positive naive T cells and/or CD4-positive central memory T cells, but also by a high bulk yield.
1. A method of producing CD4-positive naïve T cells and/or CD4-positive central memory T cells expressing CD62L and/or CCR7, the method comprising the following steps:
(1) removing CD8-positive T cells or collecting CD4-positive T cells from a cell population containing T cells; and
(2) culturing the cell population obtained by the step (1) in the presence of the following (A) and (B), and for the whole period or a partial period during the culture and/or during expansion after the culture, culturing the cell population in the presence of the following (C):
(A) at least one polypeptide selected from the group consisting of fibronectin, fibronectin fragments and their mixtures,
(B) a CD3 ligand, or a CD3 ligand and a CD28 ligand,
(C) interleukin-4 in an amount of 0.01 to 100 ng/ml.
2. The method according to claim 1 , wherein the step (2) is a step of culturing the cell population in the presence of (A) and (B) and in the presence of (C), and then expanding the obtained cell population in the presence of (C).
3. The method according to claim 1 , wherein the step (2) is a step of culturing the cell population in the presence of (A) and (B) and in the absence of (C), and then expanding the obtained cell population in the presence of (C).
4. The method according to claim 1 , wherein the step (2) is a step of culturing the cell population in the presence of (A) and (B) and in the presence of (C), and then expanding the obtained cell population in the absence of (C).
5. The method according to claim 1 , further comprising a step for foreign gene transfer.
6. The method according to claim 1 , wherein the cell population containing T cells in step (1) is peripheral blood mononuclear cells (PBMC).