2004 Fiscal Year Final Research Report Summary
3 dimensional fat regeneration using Adipose derived stem cells
| Project/Area Number |
14571723
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Plastic surgery
|
|---|
| Research Institution | Nippon Medical School |
|---|
Principal Investigator |
MIZUNO Hiroshi Nippon Medical School, Department of Plastic and Reconstructive Surgery, Assistant Professor, 医学部, 講師 (80343606)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
IGOTA Shinichi Nippon Medical School, Department of Plastic and Reconstructive Surgery, Instructor, 医学部, 助手 (30339361)
MURAKAMI Masahiro Nippon Medical School, Department of Plastic and Reconstructive Surgery, Associate Professor, 医学部, 講師 (00239500)
HYAKUSOKU Hiko Nippon Medical School, Department of Plastic and Reconstructive Surgery, Professor, 大学院・医学研究科, 教授 (00165135)
|
|---|
| Project Period (FY) |
2002 – 2004
|
| Keywords | Mesenchymal stem cells / Fat / Regenerative Medicine / Tissue Engineering |
|---|
| Research Abstract |
The species of adipose derived stem cells was changed from human to GFP transgenic mice in this project because human specimen was relatively difficult to obtain. Accordingly, we firstly identified that similar population of human adipose derived stem cells(ASC) can be isolated from GFP transgenic mice.
After the proliferation of ASC in control medium, they were induced into adipogenic differentiation in the induction medium for 1 week, followed by implanted subcutaneously into athymic mice with vehicles including non-woven polyglycolic acid, type I collagen sponge, hyaluronic acid and fibrin glue for in vivo adipogenesis. After 4 and 8 weeks of implantation, these specimen were evaluated macroscopically, histologically and immunohistochemically.
After 4 weeks of implantation, ASC with fibrin glue revealed macroscopically half transparent with small angiogenesis. Furthermore, after 8 weeks of implantation, this specimen revealed yellish in color with rounder shape. In addition, these specimens strongly emitted green fluorescence through fluorescent microscope, which suggested that transplanted ASC contributed to in vivo adipogenesis. These findings were also confirmed by immunnohistochemical analysis using anti GFP monoclonal antibodies. Both H&E and Oil Red 0 staining revealed that 3 dimensional adipose tissue which contains small size of lipid droplets in each cell were engineered.
In addition to fibrin glue, ASC with type I collagen sponge also contributed to in vivo adipogenesis, which may be another suitable scaffold for adipose tissue engineering. On the other hand, we didn't confirm any adipogenesis using other scaffolds. The gene expression of PPARγ using RT-PCR is currently being investigated.
Finally, real-time PCR analysis revealed that ASC derived from female GFP mice has a 1.54 fold greater potential of gene expression of PPARγ compared with that from male mice.
|
