| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
a Division of Developmental Biology, Department of Parasitology,
b Third Department of Internal Medicine, and
c Second Department of Anatomy, Nara Medical University, Kashihara, Nara, Japan;
d Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan
Key Words. Embryonic stem cell • Dithizone • Pancreatic differentiation • Insulin-secreting cells • Cell transplantation
Masahide Yoshikawa, M.D., Division of Developmental Biology, Department of Parasitology, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8521, Japan. Telephone: 81-744-22-3051, ext 2250; Fax: 81-744-24-7122; e-mail: myoshika{at}nmu-gw.naramed-u.ac.jp
Background and Aims. Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro. We have recently identified the emergence of cellular clusters within differentiated ES cell cultures by staining with dithizone (DTZ). DTZ is a zinc-chelating agent known to selectively stain pancreatic beta cells because of their high zinc content. The aim of the present study was to investigate the characteristics of DTZ-stained cellular clusters originating from ES cells.
Methods. Embryoid bodies (EBs), formed by a 5-day hanging drop culture of ES cells, were allowed to form outgrowths in the culture. The outgrowths were incubated in DTZ solution (final concentration, 100 µg/ml ) for 15 minutes before being examined microscopically. The gene expression of endocrine pancreatic markers was also analyzed by reverse transcriptase-polymerase chain reaction. In addition, insulin production was examined immunohistochemically, and its secretion was examined using enzyme-linked immunosorbent assay.
Results. DTZ-stained cellular clusters appeared after approximately 16 days in the EB culture and became more apparent by day 23. They were found to be immunoreactive to insulin and expressed pancreatic-duodenal homeobox 1 (PDX1), proinsulin 1, proinsulin 2, glucagon, pancreatic polypeptide, glucose transporter-2 (GLUT2), and islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) mRNA. They were also able to secrete detectable amounts of insulin.
Conclusions. ES cell-derived DTZ-positive cellular clusters possess characteristics of the endocrine pancreas, including insulin secretion. Further, DTZ staining is a useful method for the identification of differentiated pancreatic islets developed from EBs in vitro.
This article has been cited by other articles:
|
|
 |
|
  J. Jo, M. Y. Choi, and D.-S. Koh Size Distribution of Mouse Langerhans Islets Biophys. J., October 15, 2007; 93(8): 2655 - 2666. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. K. Mfopou, V. De Groote, X. Xu, H. Heimberg, and L. Bouwens Sonic Hedgehog and Other Soluble Factors from Differentiating Embryoid Bodies Inhibit Pancreas Development Stem Cells, May 1, 2007; 25(5): 1156 - 1165. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Okuno, K. Minami, A. Okumachi, K. Miyawaki, N. Yokoi, S. Toyokuni, and S. Seino Generation of insulin-secreting cells from pancreatic acinar cells of animal models of type 1 diabetes Am J Physiol Endocrinol Metab, January 1, 2007; 292(1): E158 - E165. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. B. Herrera, S. Bruno, S. Buttiglieri, C. Tetta, S. Gatti, M. C. Deregibus, B. Bussolati, and G. Camussi Isolation and Characterization of a Stem Cell Population from Adult Human Liver Stem Cells, December 1, 2006; 24(12): 2840 - 2850. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Tayaramma, B. Ma, M. Rohde, and H. Mayer Chromatin-Remodeling Factors Allow Differentiation of Bone Marrow Cells into Insulin-Producing Cells Stem Cells, December 1, 2006; 24(12): 2858 - 2867. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. S. Narang and R. I. Mahato Biological and biomaterial approaches for improved islet transplantation. Pharmacol. Rev., June 1, 2006; 58(2): 194 - 243. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Minami, M. Okuno, K. Miyawaki, A. Okumachi, K. Ishizaki, K. Oyama, M. Kawaguchi, N. Ishizuka, T. Iwanaga, and S. Seino Lineage tracing and characterization of insulin-secreting cells generated from adult pancreatic acinar cells PNAS, October 18, 2005; 102(42): 15116 - 15121. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. K.C. Brolen, N. Heins, J. Edsbagge, and H. Semb Signals From the Embryonic Mouse Pancreas Induce Differentiation of Human Embryonic Stem Cells Into Insulin-Producing {beta}-Cell-Like Cells Diabetes, October 1, 2005; 54(10): 2867 - 2874. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Shi, L. Hou, F. Tang, W. Jiang, P. Wang, M. Ding, and H. Deng Inducing Embryonic Stem Cells to Differentiate into Pancreatic {beta} Cells by a Novel Three-Step Approach with Activin A and All-Trans Retinoic Acid Stem Cells, May 1, 2005; 23(5): 656 - 662. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Wobus and K. R. Boheler Embryonic Stem Cells: Prospects for Developmental Biology and Cell Therapy Physiol Rev, April 1, 2005; 85(2): 635 - 678. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. J Burns, S. J Persaud, and P. M Jones Stem cell therapy for diabetes: do we need to make beta cells? J. Endocrinol., December 1, 2004; 183(3): 437 - 443. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. T. Ku, N. Zhang, A. Kubo, R. O'Connor, M. Mao, G. Keller, and J. S. Bromberg Committing Embryonic Stem Cells to Early Endocrine Pancreas In Vitro Stem Cells, December 1, 2004; 22(7): 1205 - 1217. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Hansson, A. Tonning, U. Frandsen, A. Petri, J. Rajagopal, M. C.O. Englund, R. S. Heller, J. Hakansson, J. Fleckner, H. N. Skold, et al. Artifactual Insulin Release From Differentiated Embryonic Stem Cells Diabetes, October 1, 2004; 53(10): 2603 - 2609. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. T. Clark, M. S. Bodnar, M. Fox, R. T. Rodriquez, M. J. Abeyta, M. T. Firpo, and R. A. R. Pera Spontaneous differentiation of germ cells from human embryonic stem cells in vitro Hum. Mol. Genet., April 1, 2004; 13(7): 727 - 739. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Nishimura, M. Yoshikawa, S. Kanda, M. Nonaka, H. Yokota, A. Shiroi, H. Nakase, H. Hirabayashi, Y. Ouji, J.-I. Birumachi, et al. Potential Use of Embryonic Stem Cells for the Treatment of Mouse Parkinsonian Models: Improved Behavior by Transplantation of In Vitro Differentiated Dopaminergic Neurons from Embryonic Stem Cells Stem Cells, March 1, 2003; 21(2): 171 - 180. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Rajagopal, W. J. Anderson, S. Kume, O. I. Martinez, and D. A. Melton Insulin Staining of ES Cell Progeny from Insulin Uptake Science, January 17, 2003; 299(5605): 363 - 363. [Full Text] [PDF]
|
|
|
|
|
|
Y. Hori, I. C. Rulifson, B. C. Tsai, J. J. Heit, J. D. Cahoy, and S. K. Kim Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells PNAS, December 10, 2002; 99(25): 16105 - 16110. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| STEM CELLS | THE ONCOLOGIST | CME | ALPHAMED PRESS JOURNALS |
