Home › Immunology › Using Human Induced Pluripotent Stem Cells for the Generation of Tumor Antigen-specific T Cells
Immunology JoVE (Open Access) Citable · DOI

Using Human Induced Pluripotent Stem Cells for the Generation of Tumor Antigen-specific T Cells

DOI: 10.3791/59997-v
What you'll learn
  • ✓ Culture human iPSCs on mouse embryonic fibroblast feeders
  • ✓ Prepare OP9/DLL1 co-culture system for T cell differentiation
  • ✓ Differentiate hiPSCs into functional CD8αβ+ T cells in vitro
  • ✓ Characterize hematopoietic progenitor cells from iPSC-derived cultures
Protocol

This article describes a method to generate functional tumor antigen-specific induced pluripotent stem cell-derived CD8αβ+ single positive T cells using OP9/DLL1 co-culture system.

Difficulty
advanced
Total time
~4–6 weeks (iPSC maintenance: ~1 week; differentiation protocol: ~3–5 weeks)
Model organism
Human induced pluripotent stem cells (hiPSCs); OP9 stromal cells (mouse); DLL1 co-culture system
Biosafety
BSL-1

Steps

1
Passage human iPSCs on mouse embryonic fibroblasts

Maintain hiPSC cultures on MEF feeders with routine passaging to preserve pluripotency and viability prior to differentiation experiments.

▶ 00:58
2
Prepare OP9/DLL1 cells for hiPSC co-culture

Establish and condition OP9 stromal cells expressing DLL1 (Delta-like ligand 1) to support T cell lineage differentiation from iPSCs.

▶ 02:30
3
Differentiate hiPSCs into CD8αβ+ T cells

Culture hiPSCs on OP9/DLL1 feeders using defined differentiation protocols to generate functional, tumor antigen-specific CD8αβ+ single-positive T cells.

▶ 03:51
4
Characterize hiPSC-derived hematopoietic progenitor cells

Analyze and quantify hematopoietic progenitor cell populations generated during the differentiation process using flow cytometry and functional assays.

▶ 08:38
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