Primary culture of proximal tubular cells (PTC) from normal mouse kidney as an in vitro model to study mechanisms of development of tubulointerstitial nephritis
| Project/Area Number |
05670944
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Kidney internal medicine
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| Research Institution | CHIBA UNIVERSITY |
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Principal Investigator |
UEDA Shiro CHIBA UNIVERSITY,SCHOOL OF MEDICINE,LECTURER, 医学部, 講師 (50201348)
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| Co-Investigator(Kenkyū-buntansha) |
HORI Junro CHIBA UNIVERSITY,SCHOOL OF MEDICIN,FELLOW, 医学部・附属病院, 医員
OGAWA Makoto CHIBA UNIVERSITY,CHIBA UNIVERSITY HOSPITAL,ASSISTANT, 医学部・附属病院, 助手 (50241956)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Tubular basement membrane (TBM) anrigen / Cell culture / Proximal tubular cells (PTC) / Tubulointerstitial nephritis / Intercellular adhesion molecule-1 (ICAM-1) / 近位尿細管上皮細胞 / ドーム形成 / ICAM-1 |
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| Research Abstract |
Cultured murine PTC clearly expressed ICAM-1 under the influence of TBM-primed T lymphocytes. These lymphocytes also demonstrated a high cytotoxic activity against PTC.Purified PTC from BALB/c mice was primary-cultured. PTC was prepared by a modified method of Bogaard et al (Biochem Pharmacol 39 : 13351345,1990). Briefly, kidney was perfused with buffer containing 0.08% (w/v) collagenase. Then, the cortical tissue was filtered through a nylon-gauze. Viable PTC was separated from other materials by isopycnic centrifugation on a discontinuous Nycodenz gradient. The confluent monolayr of PTC showed typical epithelial morphology with cobblestone-like appearanc. Typical dome formation was observed in PTC cultures. These cells also expressed gamma-GTP activity. TBM-primed nylon-column non-adherent lymphocytes were prepared from BALB/c mice immunized with mouse TBM Ag in adjuvant.
Cocultured PTC with TBM-primed lymphocytes induced ICAM-1 expression in PTC.The lymphocytes also had a strong cytotoxic activity without complement. Depletion of T cells from these lymphocytes removed the cytotoxic activity. Neither liver antigen-primed nor DNP-primed lymphocytes had any cytotoxicity against PTC.
This simple syngeneic model may allow further molecularbiological examination of tubulointerstitial nephritis.
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Report
(3 results)
Research Products
(9 results)
