Toward the establishment of culture system to restore in vivo phenotypes of podocytes

2014 Fiscal Year Final Research Report

• Project/Area Number: 24591190
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Kidney internal medicine
• Research Institution: Niigata University
• Principal Investigator: YAOITA Eishin 新潟大学, 医歯学系, 准教授 (00157950)
• Co-Investigator(Kenkyū-buntansha): YOSHIDA Yutaka 新潟大学, 医歯学系, 講師 (40182795) ; TAKEUCHI Kousei 愛知医科大学, 医学部, 教授 (90206946) ; YAMAMOTO Tadashi 新潟大学, 医歯学系, 教授 (30092737)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: ポドサイト / 培養 / 継代 / トリプシン / ヘパリン / ビタミンA / 細胞外基質 / 細胞密度

Outline of Final Research Achievements

Glomerular epithelial cells in the kidney (podocytes) play critical roles in maintenance of glomerular structure and glomerular ultrafiltration. Establishment of culture system remaining in vivo phenotypes of podocytes is essential for studies of podocytes, although podocytes drastically decrease podocyte-specific gene expressions with time in the primary culture. Of these, decrease in nephrin was the most evident. In this study, we evaluated effect of various culture conditions on the gene expression in primary culture of podocytes to improve phenotypes of cultured cells to restore in vivo phenotypes. Consequently, subculture on day 3 of culture of isolated glomeruli, detachment with non-enzymatic solution, addition of heparin, decrease in fetal bovine serum, adequate cell density, and extracellular matrices enable expression of podocyte-specific genes at the level of in vivo. In addition, dexamethasone and all trans-retinoic acid induce cellular processes of cultured podocytes.

Free Research Field

腎臓病理学