| Project/Area Number |
13680925
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | University of Tsukuba |
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Principal Investigator |
HATTORI Kazunori Univ. of Tsukuba, Urology, Assist. Prof., 臨床医学系, 講師 (50312848)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAI Koji Univ of Tsukuba, Urology, Assist. Prof., 臨床医学系, 講師 (90272195)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2001: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | ureter / bio-material / collagen / hybrid organ / 人工尿路 / 再生医療 |
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| Research Abstract |
1. Primary culture of pocine uroepitheial cells and stromal cells.
Primary culture of pocine urothelial cells and stromal cells was established from ureteral tissues. A urothelial sheet was peeled off the submucosa with aid of a dissecting microscope. The epithelial sheet and muscular tissues were cut into 1-2mm fragments, and digested with dispase or collagenase. Both epithelial cells and stromal cells could be propagated several times.
2. Construction of tissue-engineered ureteral tissue with collagen gel.
Ureteral stromal cells were embedded into type 1 collagen gel, and put into a glass mold. After a week, ureteral epithelial cells were seeded inside of the collagen gel tube. However, because the gel was significantly shrunken and became fragile during culture period, it was impossible to seed epithelial cells into the lumen of the collagen tube. Further modification will be needed to construct ureteral tissues with collagen gel in vitro.
3. Preparation of Ureteral Acellular Matrix Graft (UAMG)
Pocine ureters were processed by treatment with PBS, Dnase and sodium deoxycholate. Acellularity of UAMG was confirmed by histological analysis. Uretral epithelial cells and stromal cells were seeded on the luminal or outer surface of the UAMG. Although both cells were attached well on the UAMG, infiltration of the cells into the deep layer of UAMG was quite limited.
4. Replacement of ureter with UAMG
Under general anesthesia, mid-portion of a pocine ureter was removed and replaced by UAMG. Four weeks later, the kidney and upper ureter showed significant hydronephroureter due to severe stenosis at the anastomosis. Histologically, normal epithelial layer or muscle layer was not observed. Instead, infiltration of mononuclear cells and myofibroblasts was observed.
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