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TISSUE-SPECIFIC STEM CELLS |
aCornea Center, Tokyo Dental College, Ichikawa, Chiba, Japan;
bDepartment of Ophthalmology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan;
cDepartment of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan;
dDepartment of Regenerative Medicine and Advanced Cardiac Therapeutics, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan
Key Words. Corneal stroma • Keratocyte • Stem cells • Bone marrow cells • Neural crest
Correspondence: Shigeto Shimmura, M.D., Department of Ophthalmology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. Telephone: +81-3-3353-1211; Fax: +81-3-3359-8302; e-mail: shige{at}sc.itc.keio.ac.jp; or Hideyuki Okano, M.D., Ph.D., Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. Telephone: +81-3-3353-1211; Fax: +81-3-3357-5445; e-mail: hidokano{at}sc.itc.keio.ac.jp
Received on March 16, 2006;
accepted for publication on July 24, 2006.
First published online in STEM CELLS EXPRESS August 3, 2006.
We report the presence of neural crest-derived corneal precursors (COPs) that initiate spheres by clonal expansion from a single cell. COPs expressed the stem cell markers nestin, Notch1, Musashi-1, and ABCG2 and showed the side population cell phenotype. COPs were multipotent with the ability to differentiate into adipocytes, chondrocytes, as well as neural cells, as shown by the expression of ß-III-tubulin, glial fibrillary acidic protein, and neurofilament-M. COP spheres prepared from E/nestin-enhanced green fluorescent protein (EGFP) mice showed induction of EGFP expression that was not originally observed in the cornea, indicating activation of the neural-specific nestin second intronic enhancer in culture. COPs were Sca-1+, CD34+, CD45–, and c-kit–. Numerous GFP+ cells were observed in the corneas of mice transplanted with whole bone marrow of transgenic mice ubiquitously expressing GFP; however, no GFP+ COP spheres were initiated from these mice. On the other hand, COP spheres from transgenic mice encoding P0-Cre/Floxed-EGFP as well as Wnt1-Cre/Floxed-EGFP were GFP+, indicating the neural crest origin of COPs, which was confirmed by the expression of the embryonic neural crest markers Twist, Snail, Slug, and Sox9. Taken together, these data indicate the existence of neural crest-derived, multipotent stem cells in the adult cornea.
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