alexa Murine Insulinoma Cell-Conditioned Medium with and#914;ETA2/Neurod1 Transduction Efficiently Induces the Differentiation of Adipose-Derived Mesenchymal Stem Cells into and#946;-Like Cells both <em>In Vitro</em> and <em>In Vivo</em> | Abstract
ISSN: 2157-7633

Journal of Stem Cell Research & Therapy
Open Access

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Research Article

Murine Insulinoma Cell-Conditioned Medium with ΒETA2/Neurod1 Transduction Efficiently Induces the Differentiation of Adipose-Derived Mesenchymal Stem Cells into β-Like Cells both In Vitro and In Vivo

Koichi Kawamoto1#, Shigeharu Yabe2#, Masamitsu Konno3#, Hideshi Ishii3, Naohiro Nishida3, Jun Koseki3, Satsuki Fukuda2, Yoshito Tomimaru1, Naoki Hama1, Hiroshi Wada1, Shogo Kobayashi1, Hidetoshi Eguchi1, Masahiro Tanemura4, Toshinori Ito5, Eun Young Lee6, Eri Mukai6, Takashi Miki6, Yuichiro Doki1, Masaki Mori1, Tatsuo S Hamazaki2, Hiroaki Nagano1* and Hitoshi Okochi2*

1Department of Surgery, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan

2Department of Regenerative Medicine, Research Institute, National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjuku-ku, Tokyo 162-8655, Japan

3Department of Frontier Science for Cancer and Chemotherapy, National Hospital Organization Kure Medical Center, Hiroshima, 737-0023, Japan

4Department of Surgery and Institute for Clinical Research, National Hospital Organization Kure Medical Center, Hiroshima, 737-0023, Japan

5Department of Complementary and Alternative Medicine, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan

6Department of Medical Physiology, Chiba University, Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan

#These authors contributed equally to the work

*Corresponding Authors:
Hiroaki Nagano, MD, PhD
Associate Professor, Department of Surgery
Osaka University, Graduate School of Medicine
Suita, Yamadaoka 2-2, Osaka 565-0871, Japan
Tel: +81-(0)6-6879-3251
Fax: +81-(0)6-6879-3259
E-mail: [email protected]
 
Hitoshi Okochi, MD, PhD
Professor, Department of Regenerative Medicine
Research Institute, National Center for Global Health and Medicine
1-21-1 Toyama, Shinjuku-ku, Tokyo 162-8655, Japan
Tel: +81-(0)3-3202-7181-2727
Fax: +81- 03-3207-1038
E-mail: [email protected]

Received date: July 10, 2014; Accepted date: July 26, 2014; Published date: July 28, 2014

Citation: Kawamoto K, Yabe S, Konno M, Ishii H, Nishida N, et al. (2014) Murine Insulinoma Cell-Conditioned Medium with BETA2/Neurod1 Transduction Efficiently Induces the Differentiation of Adipose-Derived Mesenchymal Stem Cells into β-Like Cells both In Vitro and In Vivo. J Stem Cell Res Ther 4:221. doi:10.4172/2157-7633.1000221

Copyright: © 2014 Kawamoto K, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Background: Mesenchymal stem cells (MSCs), including adipose tissue-derived mesenchymal stem cells (ADSCs), are multipotent and can differentiate into various cell types, including pancreatic β cells. Therefore, ADSCs present a potential cell source for the treatment of type 1 diabetes mellitus (T1DM). However, current in vitro protocols are insufficient to induce fully matured insulin-producing β cells. In this study, we assessed the effectiveness of overexpression of ΒETA2 (NeuroD1), a member of the basic helix–loop–helix transcription factor family, with murine insulinoma cell line-derived conditioned medium (MIN6-CM) to improve the differentiation capacity of ADSCs into insulin-producing cells.
Method: Murine ADSCs were isolated from C57BL/6 mice, transduced with several transcriptional factors (TFs), and stable transfectants were established. MIN6-CM was prepared. Syngeneic recipient mice were rendered diabetic by a single injection of streptozotocin, and differentiated cells were transplanted under the kidney capsule of recipient mice. Next, blood glucose levels were monitored.
Results: CM alone was sufficient to induce insulin mRNA expression in vitro. However, other TFs were not detected. ADSCs cultured with MIN6-CM induced insulin expressions in vitro, but other β cell-related TFs were been detected. However, BETA2 transduction in MIN6-CM resulted in robust expression of multiple β cell phenotypic markers. Moreover, insulin content analysis revealed insulin protein expression in vitro. Furthermore, in vivo transplant studies revealed the effectiveness of the simultaneous use of BETA2 transduction with the CM.
Conclusion: These results suggest that the balance of cytokines and growth factors in addition to gene manipulation would benefit the efficient differentiation of ADSCs into pancreatic β cells. Our technology could provide a path to β cell differentiation and novel cell replacement-based therapies for T1DM.

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