Protocol to generate human T cell progenitors that repopulate the thymus

Elena S Philonenko1, Baoyun Zhang1, Eugene Albert2

  • 1CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.

STAR Protocols
|August 7, 2026
PubMed

Human pluripotent stem cells (hPSCs) provide a powerful platform for generating T cell progenitors and investigating hematopoietic stem cell (HSC)-independent T cell development. Here, we present a protocol for the directed differentiation of hPSCs to generate early progenitors of human T cells that repopulate the thymus long term and gradually develop into mature T cells in vivo. The protocol includes primary hematopoietic differentiation of hPSCs in defined culture conditions, a secondary differentiation step to prime T lymphopoiesis, and mouse transplantation procedures. For complete details on the use and execution of this protocol, please refer to Philonenko et al.1.

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