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First published online November 8, 2007
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2007-0348v1
26/3/706    most recent
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Submitted on May 8, 2007
Accepted on October 25, 2007

EMBRYONIC STEM CELLS

Direct Development of Functionally Mature Tryptase/chymase Double Positive Connective Tissue-Type Mast Cells from Primate ES Cells

Feng Ma 1, Naotomo Kambe 2, Dan Wang 3, Gen Shinoda 4, Hisanori Fujino 4, Katsutsugu Umeda 4, Akihiro Fujisawa 2, Lina Ma 4, Hirofumi Suemori 5, Norio Nakatsuji 5, Yoshiki Miyachi 2, Ryuzo Torii 6, Kohichiro Tsuji 3, Toshio Heike 4, Tatsutoshi Nakahata 4*

1 Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto 606-8507, Japan; Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
2 Department of Dermatology, Kyoto University Graduate School of Medicine, Kyoto 606-8507, Japan
3 Division of Cellular Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan
4 Department of Pediatrics, Kyoto University Graduate School of Medicine, Kyoto 606-8507, Japan
5 Research Center for Regenerative Medicine, Kyoto University, Kyoto 606-8507, Japan
6 Research Center for Animal Life Science, Shiga University of Medical Science, Ohtsu 520-2192, Japan

* To whom correspondence should be addressed. E-mail: tnakaha{at}kuhp.kyoto-u.ac.jp.


  Abstract

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 co-cultured with the murine aorta-gonad-mesonephros-derived stromal cell line AGM-S1 differentiated into cobblestone (CS)-like cells by day 10 to 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 three 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 co-expressed a high affinity IgE receptor on their surface, which was up-regulated 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.

______________________________________________________________________________

F.M. and N.K. contributed equally to this study.

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






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