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First published online November 8, 2007
Stem Cells Vol. 26 No. 3 March 2008, pp. 706 -714
doi:10.1634/stemcells.2007-0348; www.StemCells.com
© 2008 AlphaMed Press

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EMBRYONIC STEM CELLS

Direct Development of Functionally Mature Tryptase/Chymase Double-Positive Connective Tissue-Type Mast Cells from Primate Embryonic Stem Cells

Feng Maa,b, Naotomo Kambef, Dan Wangb, Gen Shinodaa, Hisanori Fujinoa, Katsutsugu Umedaa, Akihiro Fujisawac, Lina Maa, Hirofumi Suemorid, Norio Nakatsujid, Yoshiki Miyachic, Ryuzo Toriie, Kohichiro Tsujib, Toshio Heikea, Tatsutoshi Nakahataa

Departments of aPediatrics and
cDermatology, Kyoto University Graduate School of Medicine, Kyoto, Japan;
bDivision of Cellular Therapy, Institute of Medical Science, University of Tokyo, Tokyo, Japan;
dResearch Center for Regenerative Medicine, Kyoto University, Kyoto, Japan;
eResearch Center for Animal Life Science, Shiga University of Medical Science, Ohtsu, Japan;
fDepartment of Dermatology, Chiba University Graduate School of Medicine, Chiba, Japan

Key Words. Embryonic stem cells • Mast cells • Primate • Development • Chymase • Tryptase

Correspondence: Tatsutoshi Nakahata, M.D., Ph.D., Department of Pediatrics, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Telephone: 81-75-751-3290; Fax: 81-75-752-2361; e-mail: tnakaha{at}kuhp.kyoto-u.ac.jp

Received May 8, 2007; accepted for publication October 25, 2007.
First published online in STEM CELLS EXPRESS   November 8, 2007.



Conditions that influence the selective development or recruitment of connective tissue-type and mucosal-type mast cells (MCs) are not well understood. Here, we report that cynomolgus monkey embryonic stem (ES) cells cocultured with the murine aorta-gonad-mesonephros-derived stromal cell line AGM-S1 differentiated into cobblestone (CS)-like cells by day 10–15. When replated onto fresh AGM-S1 with the addition of stem cell factor, interleukin-6, and Flt3 ligand, these CS-like cells displayed robust growth and generated almost 100% tryptase/chymase double-positive MCs within 3 weeks. At all time points, the percentage of tryptase-positive cells did not exceed that of chymase-positive cells. These ES-derived MCs were CD45+/Kit+/CD31+/CD203c+/HLA-DR– and coexpressed a high-affinity IgE receptor on their surface, which was upregulated after IgE exposure. Electron microscopy showed that they contained many electron dense granules. Moreover, ES-derived MCs responded to stimulation by via IgE and substance P by releasing histamine. These results indicate that ES-derived MCs have the phenotype of functionally mature connective tissue-type MCs. The rapid maturation of ES-derived MCs suggests a unique embryonic pathway in primates for early development of connective tissue-type MCs, which may be independent from the developmental pathway of mucosal-type MCs.

Disclosure of potential conflicts of interest is found at the end of this article.






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