Neural Crest–Derived Stem Cells Migrate and Differentiate
Into Cardiomyocytes After Myocardial Infarction
Yuichi Tamura, Keisuke Matsumura, Motoaki Sano, Hidenori Tabata, Kensuke Kimura, Masaki Ieda,
Takahide Arai, Yohei Ohno, Hideaki Kanazawa, Shinsuke Yuasa, Ruri Kaneda, Shinji Makino,
Kazunori Nakajima, Hideyuki Okano, Keiichi Fukuda
Objective—We recently demonstrated that primitive neural crest–derived (NC) cells migrate from the cardiac neural crest
during embryonic development and remain in the heart as dormant stem cells, with the capacity to differentiate into
various cell types, including cardiomyocytes. Here, we examined the migration and differentiation potential of these
cells on myocardial infarction (MI).
Methods and Results—We obtained double-transgenic mice by crossing protein-0 promoter-Cre mice with Floxed–enhanced
green fluorescent protein mice, in which the NC cells express enhanced green fluorescent protein. In the neonatal heart, NC
stem cells (NCSCs) were localized predominantly in the outflow tract, but they were also distributed in a gradient from base
to apex throughout the ventricular myocardium. Time-lapse video analysis revealed that the NCSCs were migratory. Some
NCSCs persisted in the adult heart. On MI, NCSCs accumulated at the ischemic border zone area (BZA), which expresses
monocyte chemoattractant protein-1 (MCP-1). Ex vivo cell migration assays demonstrated that MCP-1 induced NCSC
migration and that this chemotactic effect was significantly depressed by an anti-MCP-1 antibody. Small NC cardiomyocytes
first appeared in the BZA 2 weeks post-MI and gradually increased in number thereafter.
Conclusion—These results suggested that NCSCs migrate into the BZA via MCP-1/CCR2 signaling and contribute to the
provision of cardiomyocytes for cardiac regeneration after MI. (Arterioscler Thromb Vasc Biol. 2011;31:582-589.)
Key Words: biology, developmental ? cytokines ? ischemic heart disease ? molecular biology
? cardiac regeneration
positive, and side-population cells.1–4All of these can differ-
entiate into various cell types, including cardiomyocytes.
However, the developmental origins of these cardiac stem
cells remain unclear.
Neural crest–derived (NC) cells constitute the fourth germ
cell layer in the developing embryo. Cells from the neural
crest are characterized by extensive migration and can differ-
entiate in various developing tissues into neurons, glial cells,
endocrine organs, bone, tooth, cartilage, pigmented epithe-
lium, cornea, and aortic smooth muscle cells.5–7In addition,
some NC cells are retained in the adult tissues as dormant
stem cells, referred to as NC stem cells (NCSCs).8–10
We recently isolated the side-population cells from mam-
malian heart.11These cells formed a neurosphere-like struc-
ture that expressed nestin and Musashi-1, which is a marker
of undifferentiated neural precursor cells12and could differ-
entiate into neurons, glial cells, smooth muscle cells, and
everal types of stem cells have been identified in adult
heart, including c-Kit-positive, Sca-1-positive, Isl-1-
cardiomyocytes. Furthermore, when these sphere-forming
cells were transplanted into the neural crest of a chick
embryo, they migrated and differentiated into neurons and
glial cells at the sympathetic ganglia, dorsal root ganglia, and
spinal nerves. They also migrated into the developing heart
region via the lateral pathway.11These results suggested the
existence of a pool of NCSCs that are dormant in the adult
myocardium. The regenerative potential of such a cell popu-
lation in injured heart remains to be determined.
This study traced the fate of NC cells in heart during
postneonatal growth and after myocardial infarction (MI)
using double-transgenic (Tg) mice harboring protein-0 (P0)
promoter-Cre13and Floxed–enhanced green fluorescent pro-
tein (EGFP).14The P0 glycoprotein was originally identified
as a Schwann cell–specific myelin protein,15but it is also
expressed in migrating neural crest cells during the early
embryonic period in chicks.16In the double-Tg mice, tran-
sient activation of the P0 promoter induces Cre-mediated
recombination, which tags subsets of the NC cells with
Received on: August 16, 2010; final version accepted on: December 9, 2010.
From the Departments of Cardiology (Y.T., K.M., M.S., K.K., M.I., T.A., Y.O., H.K., S.Y., R.K., S.M., K.F.), Anatomy (H.T., K.N.), and Physiology
(H.O.), Keio University School of Medicine, Tokyo, Japan.
Drs Tamura and Matsumura contributed equally to this work.
Correspondence to Motoaki Sano, Department of Cardiology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo 160-8582,
Japan (E-mail email@example.com); or Keiichi Fukuda, Department of Cardiology, Keio University School of Medicine, 35 Shinanomachi
Shinjuku-ku, Tokyo 160-8582, Japan (E-mail firstname.lastname@example.org).
© 2011 American Heart Association, Inc.
Arterioscler Thromb Vasc Biol is available at http://atvb.ahajournals.orgDOI: 10.1161/ATVBAHA.110.214726
Biomedical Innovation, Japan, and by research grants from the
Ministry of Education, Science and Culture, Japan.
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Tamura et alNeural Crest Cells and Cardiac Repair