| Budget Amount *help |
¥2,548,000 (Direct Cost: ¥1,960,000、Indirect Cost: ¥588,000)
Fiscal Year 2010: ¥1,235,000 (Direct Cost: ¥950,000、Indirect Cost: ¥285,000)
Fiscal Year 2009: ¥1,313,000 (Direct Cost: ¥1,010,000、Indirect Cost: ¥303,000)
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| Research Abstract |
Scar fibroblasts (SFs) were isolated from ferret vocal folds electrocauterized two weeks previously (N=2). Adipose-derived stem cells (ASCs) were isolated from ferret lipoaspirated subcutaneous abdominal fat (N=2). For co-culture experiments, the two cell types were combined in Transwell plates for 6 days, followed by 1 or 3 days of monoculture after removing the upper chamber. Assays were then performed on cells and media from the bottom chamber. We measured (1) the production of hyaluronic acid (HA), collagen and hepatocite growth factor (HGF) via enzyme-linked immunosorbent assays (ELISA) (2) the expression of α-smooth muscle actin (α-SMA), (3) cell proliferation and (4) apoptosis of SFs (2, 3 & 4 via flow cytometry). Other experiments examined the effects of HGF on SFs and the effects of HGF neutralization in the co-culture system. We demonstrated that co-culture led to significant decreases in SF collagen production (p<.05), proliferation (p<.05), and α-SMA expression (p<.05), while HA production increased (p<.05). Co-culture also increased HGF secretion from ASCs (p<.05). Neutralization of HGF abolished the inhibitory effects of ASCs on SF collagen synthesis (p<.05). We concluded that ASCs influence SFs to adopt a less fibrotic profile. It appears that HGF is at least one of the soluble factors responsible for this effect. Implanted ASCs could potentially ameliorate vocal fold scar by acting as a long-term, intrinsic source of HGF
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