Development of an in vitro co-culture system for elucidation of host-gut microbiota symbiotsis

2020 Fiscal Year Final Research Report

• Project/Area Number: 18H02126
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 38020:Applied microbiology-related
• Research Institution: Kyoto University
• Principal Investigator: Katayama Takane 京都大学, 生命科学研究科, 教授 (70346104)
• Co-Investigator(Kenkyū-buntansha): 加藤 紀彦 京都大学, 生命科学研究科, 助教 (40724612) ; 神戸 大朋 京都大学, 生命科学研究科, 准教授 (90303875)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 共培養 / アピカル嫌気 / 嫌気性細菌 / 腸上皮細胞

Outline of Final Research Achievements

We have developed an apical anaerobic device in this study. The device was able to keep dissolved oxygen concentrations of apical side and basal side of Caco-2 monolayer at < 0.3 % and < 60 %, respectively, at least for 5 days. Transepithelial electrical resistance values were comparable to that of the cells maintained in CO2 incubator. Fluorescence immunostaining of claudin-2 revealed clear tight-junction formation of the cells. When obligate anaerobes belonging to Bacteroides and Bifidobacterium genera were added to the apical medium, the growth of these bacteria was stimulated by 10- to 1,000,000-folds. Transepithelial electrical resistance was comparable between before and after co-cultivation. These results indicate that the device is useful to examine cross-talk between intestinal epithelial cells and anaerobic microbes at the molecular levels.

Free Research Field

応用微生物学

Academic Significance and Societal Importance of the Research Achievements

本助成においてアピカル嫌気培養デバイスの有効性を示すことが出来た。本デバイスの特許出願に基づいた共同研究を国内企業2社と進めており、2022年もしくは2023年には実用化が見込まれている。本装置が汎用化されることで、よりin vivoに近い状態で共生・疾患メカニズムを解析することが可能となると考えられる。