Journal of Nippon Medical School: Vol.71 No.4 page.240

Journal of Nippon Medical School

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ArticleTitle Chondrogenic and Osteogenic Differentiation of Adipose-derived Stem Cells Isolated from GFP Transgenic Mice

AuthorList Rei Ogawa¹, Hiroshi Mizuno¹, Hiko Hyakusoku¹, Atsushi Watanabe², Makoto Migita² and Takashi Shimada²

Affiliation ¹Department of Plastic and Reconstructive Surgery, Nippon Medical School
²Department of Biochemistry and Molecular Biology, Nippon Medical School

Language EN

Volume 71

Issue 4

Year 2004

Page 240-241

Received

Accepted

Keywords

Abstract
Recent studies suggest that human adipose tissue contain pluripotent cells similar to bone marrow-derived stromal cells (BSCs)^1-3. Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice⁴, we have previously demonstrated that BSCs differentiate into a variety of cell lineages both in vitro and in vivo⁵. In the present study, we extend this approach to characterize adipose-derived stromal cells (ASCs)⁶. These cells derived from human are sometimes called processed lipoaspirate (PLA) cells. ASCs were prepared from inguinal fat pads of GFP transgenic mice after extensive washing with PBS and treatment with collagenase. After the primary culture in control medium (DMEM+10%FBS), the cells were incubated in either chondrogenic medium (DMEM+1%FBS+insulin+ascorbate 2-phosphate+TGF-beta 1) or osteogenic medium (DMEM+10%FBS+dexamethasone+ascorbate-2-phosphate+beta-glycerophosphate) for two to four weeks. Chondrogenic differentiation was assessed by Alcian blue staining, while osteogenic differentiation was by von Kossa and Alkaline phosphatase staining. ASCs incubated in chondrogenic medium induced Alcian blue positive cells. Incubation with osteogenic medium became positive for von Kossa and Alkaline phosphatase staining. No osteochondrogenic differentiation was observed in cells incubated with control medium. This cell population can be easily identified through fluorescence microscope, it should be an ideal source of ASCs for further experiments of stem cell biology and tissue engineering.

Correspondence to Rei Ogawa, MD, Department of Plastic and Reconstructive Surgery, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8603. Japan
r.ogawa@nms.ac.jp

ArticleTitle	Chondrogenic and Osteogenic Differentiation of Adipose-derived Stem Cells Isolated from GFP Transgenic Mice
AuthorList	Rei Ogawa¹, Hiroshi Mizuno¹, Hiko Hyakusoku¹, Atsushi Watanabe², Makoto Migita² and Takashi Shimada²
Affiliation	¹Department of Plastic and Reconstructive Surgery, Nippon Medical School ²Department of Biochemistry and Molecular Biology, Nippon Medical School
Language	EN
Volume	71
Issue	4
Year	2004
Page	240-241
Received
Accepted
Keywords
Abstract	Recent studies suggest that human adipose tissue contain pluripotent cells similar to bone marrow-derived stromal cells (BSCs)^1-3. Taking advantage of homogeneously marked cells from green fluorescent protein (GFP) transgenic mice⁴, we have previously demonstrated that BSCs differentiate into a variety of cell lineages both in vitro and in vivo⁵. In the present study, we extend this approach to characterize adipose-derived stromal cells (ASCs)⁶. These cells derived from human are sometimes called processed lipoaspirate (PLA) cells. ASCs were prepared from inguinal fat pads of GFP transgenic mice after extensive washing with PBS and treatment with collagenase. After the primary culture in control medium (DMEM+10%FBS), the cells were incubated in either chondrogenic medium (DMEM+1%FBS+insulin+ascorbate 2-phosphate+TGF-beta 1) or osteogenic medium (DMEM+10%FBS+dexamethasone+ascorbate-2-phosphate+beta-glycerophosphate) for two to four weeks. Chondrogenic differentiation was assessed by Alcian blue staining, while osteogenic differentiation was by von Kossa and Alkaline phosphatase staining. ASCs incubated in chondrogenic medium induced Alcian blue positive cells. Incubation with osteogenic medium became positive for von Kossa and Alkaline phosphatase staining. No osteochondrogenic differentiation was observed in cells incubated with control medium. This cell population can be easily identified through fluorescence microscope, it should be an ideal source of ASCs for further experiments of stem cell biology and tissue engineering.
Correspondence to	Rei Ogawa, MD, Department of Plastic and Reconstructive Surgery, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8603. Japan r.ogawa@nms.ac.jp