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

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.

This article has been cited by other articles:

				F. Ma, Y. Ebihara, K. Umeda, H. Sakai, S. Hanada, H. Zhang, Y. Zaike, E. Tsuchida, T. Nakahata, H. Nakauchi, et al. Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis PNAS, September 2, 2008; 105(35): 13087 - 13092. [Abstract] [Full Text] [PDF]