Thymopentin enhances the generation of T-cell lineage derived from human embryonic stem cells in vitro

• Published in: Experimental cell research 2015-02, Vol.331 (2), p.387-398
• Main Authors: Zhu, Ming-Xia, Wan, Wen-Li, Li, Hai-Shen, Wang, Jing, Chen, Gui-An, Ke, Xiao-Yan
• Format: Article
• Language: English
• Subjects: Antigens, CD34 - biosynthesis; Antigens, CD34 - metabolism; Antigens, CD7 - metabolism; Antigens, Surface - biosynthesis; Cell differentiation; Cell Lineage; Cells, Cultured; Cellular biology; Embryonic Stem Cells - cytology; Embryos; Hematopoietic progenitor cells; Hematopoietic Stem Cells - cytology; Human embryonic stem cells; Humans; Interferon-gamma - biosynthesis; Interleukin-2 - biosynthesis; Leukocyte Common Antigens - metabolism; Lymphopoiesis - drug effects; Peptides; Pluripotent Stem Cells - cytology; Stem cells; Studies; T lymphocytes; T-Lymphocytes - cytology; Thymopentin; Thymopentin - pharmacology; Tumor Necrosis Factor-alpha - biosynthesis
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2014.12.012

Thymopentin is a group of biologically active peptide secreted mainly by the epithelial cells of thymic cortex and medulla. Whether it promotes T cells production from human embryonic stem cells(hESCs) in vitro remains an elusive issue. In the present study, we develop a novel strategy that enhances T-cell lineage differentiation of hESCs in collagen matrix culture by sequential cytokine cocktails treatment combined with thymopentin stimulation. We observed that approximately 30.75% cells expressed CD34 on day 14 of the cultures and expressed the surface markers of erythroid, lymphoid and myeloid lineages. The results of colony assays and gene expressions by RT-PCR analysis also demonstrated that hematopoietic progenitor cells (HPCs) derived from hESCs were capable of multi-lineage differentiation. Further study revealed that culturing with thymopentin treatment, the CD34+CD45RA+CD7+ cells sorted from HPCs expressed T-cell-related genes, IKAROS, DNTT, TCRγ and TCRβ, and T-cell surface markers, CD3, cytoplasmic CD3, CD5, CD27, TCRγδ, CD4 and CD8. The differentiated cells produced the cytokines including IFN-γ, IL-2 and TNF-α in response to stimulation, providing the evidence for T-cell function of these cells. In conclusion, thymopentin enhances T-cell lineage differentiation from hESCs in vitro by mimicking thymus peptide environment in vivo. •Human collagen IV is sufficient to support hematopoietic differentiation from hESCs.•CD34+CD45RA+CD7+ populations commit T cells by thymopentin stimulation in vitro.•Thymopentin could simulate the polypeptide environment of thymus.