Table 2 Generation methods and characteristics of mouse and human Clec9a⁺ DCs

Source of cells

Bone marrow cells.

Peripheral blood monocytes, CD34⁺ progenitors, iPSCs.

In vivo expansion via Flt3L injection or inoculation of Flt3L-expressing B16 melanoma cells.

Culture conditions

Cultured for 9–10 days with semi-medium changes every 4 days; GM-CSF protocols for 7–10 days are less effective for Clec9a⁺ expression but may be used in combination with Flt3L to enhance specific results.

8–21 days culture; includes stages of amplification and differentiation.

Clec9a expression observed after additional culturing intervals; use of OP9 and OP9-DL1 cells in CD34⁺ progenitor differentiation; Notch signaling required for iPSC-derived DC differentiation.

Key cytokines

mFlt3L; GM-CSF can be used as an alternative but is less effective alone.

hGM-CSF, IL-4, hFlt3L, hSCF, hTPO, hIL-7.

Key cytokines support different stages: hFlt3L and hSCF crucial for DC subset differentiation; estrogen withdrawal in HoxB8-Flt3L system for timing flexibility.

Characteristics of resulting DCs

Predominantly CD8α⁺ Clec9a⁺ cDC1s; effective in antigen uptake.

CD141⁺ cDC1s and CD1c⁺ DCs; resemble blood DCs with markers like XCR1, CADM1, TLR3 (but not TLR4 or TLR9); iPSC-derived DCs homologous to peripheral blood cDC1s.

iPSC-derived DCs offer a scalable and controlled model for DC biology and therapeutic research.