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CO-CULTURE DEVICE AND CO-CULTURE METHOD FOR BACTERIUM SUCH AS ANAEROBIC BACTERIUM AND EPITHELIAL CELLS UPDATE

外国特許コード F180009620
整理番号 5450
掲載日 2018年11月20日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2017JP039206
国際公開番号 WO 2018079793
国際出願日 平成29年10月30日(2017.10.30)
国際公開日 平成30年5月3日(2018.5.3)
優先権データ
  • 特願2016-211626 (2016.10.28) JP
発明の名称 (英語) CO-CULTURE DEVICE AND CO-CULTURE METHOD FOR BACTERIUM SUCH AS ANAEROBIC BACTERIUM AND EPITHELIAL CELLS UPDATE
発明の概要(英語) To provide a means for co-culturing a bacterium such as an anaerobic bacterium and epithelial cells, disclosed is a culture system for co-culturing a first cell group consisting of one or more kinds of cells and a cell layer or tissue formed of a second cell group consisting of one or more kinds of cells different from the former cells, said culture system being characterized by comprising: a first culture tank for co-culturing under anaerobic conditions the first cell group consisting of one or more kinds of cells of, for example, an anaerobic bacterium and the cell layer or tissue formed of the second cell group consisting of one or more kinds of cells such as epithelial cells; a second culture tank for pooling a liquid culture medium under aerobic conditions; one or more matter exchange structures that are disposed so as to connect the first culture tank to the second culture tank; and the aforesaid cell layer or tissue that is disposed so as to cover the surface on the first culture tank side of the matter exchange structure(s).
従来技術、競合技術の概要(英語) BACKGROUND ART
Intestinal bacteria, human and animal intestinal bacteria that is always present while Gliocladiunz in a general term.The number and types of bacteria in the intestine is very large, in humans, about 1000 billion per fecal 1 g the presence of bacteria in the intestine and even more, at least about one degree and the type 100, also referred to as unidentified bacteria are many.From the inside of the intestine under anaerobic conditions, many of the anaerobic bacteria are bacteria in the intestine is considered.
Intestinal bacteria are, present in the gut absorption of a variety of materials, useful for human and animal intestinal metabolites can be discharged to, the digestion of food ingested in play an auxiliary role.In addition, the bacteria in the intestine, bowel extends normally in a stable configuration the intestinal flora, entering into the intestines of the human and animal pathogens to prevent the growth of the foreign, to prevent the occurrence of a disease caused by pathogens are responsible for are considered.On the other hand, the bacteria in the intestine, as its metabolites, such as carcinogens or toxic substance, an adverse effect on human and animal survival may be the production of harmful substances.These harmful substances in the intestine of the human and animal acts directly on, the absorption from the intestine as well as systemic blood flow delivered through a variety of action and cause a disorder or disease, human and animal health is impaired.
In this way, the number and types Gliocladiunz in the gut bacteria in the intestine of the various, intestinal bacteria are intestinal flora is formed, while maintaining a state of healthy human and animal remains viable and is essential for the presence of, and even the cause of a disease, in human and animal, the quality of the health of the left and right is considered to be very important are present.
On the other hand, the number and types of bacteria in the intestine, as well as the specific structure of the intestinal flora, each individual, different for each individual, in addition, the same individual, the individual, diet and the environment such as external factors such as the condition of the living environment by the change of the internal factor of, the type of bacteria in the intestine or a variation in the ratio are needed.Therefore, the intestinal flora specific arrangement of the effect on human and animal health conditions, and continue with daily changes are considered.
In recent years, such the presence of bacteria in the intestine and the intestinal flora human and animal health conditions associated with the large and the effect of the importance of attention.Then, the intestinal flora and intestinal bacteria would have an adverse effect on health and health factors contributing to elucidate the factors, human and animal health of the intestinal bacteria and, in the treatment and prevention of various diseases and the establishment of a method a very forceful momentum demand.Therefore, in order to elucidate the picture of the intestinal flora, enteric bacteria in the intestine of the human and animal from the collected and isolated culture, attempts have been made to identify the type and characteristics, can be cultured under aerobic conditions for bacteria in the intestine of the part, the analysis has been, in the presence of intestinal epithelial cells is performed by the characteristics of the confirmation test.However, intestinal bacteria, absolute anaerobic in many cases, more than half its difficult to culture, identification of bacteria and characteristics of the check is not yet in many cases.Therefore, the type of bacteria in the intestine is applied to the variation of the ratio of the external factor, also in the relationship between the factors, is accurately known is not reached.
First of all, intestinal bacteria, substantially oxygen-free absolute anaerobic conditions in the intestine based on the fact that lived in, culture dishes were placed in an oxygen-free chamber is held in an oxygen-free conditions in the intestine is reproduced, the absolute anaerobic culturing the bacteria in the intestine in the proposed method has been used.In this method, a part of the growing in culture bacteria in the intestine is effective.However, in this method, the composition of the culture liquid in the cells need to be optimized in accordance with, on the other hand, to optimize and increase only the portion of the types of intestinal bacteria, and intestinal flora other anaerobic bacteria culture of the flame retardant is grown at the same time since it is difficult, has been required to be improved.
Therefore, the bacteria in the intestine, intestinal epithelial cells of the intestinal tract by considering the fact that a viable and in contact with each other, the bacteria in the intestine, are cultured in a culture solution is simply not, in the same manner as in the intestine, intestinal epithelial cells to coexist, intestinal epithelial cells or equal to that of the cells to create a single layer film, or, cell culture insert a cell culture system used in the gastrointestinal tract on the filter of the epithelial cells or the equivalent of cells of a single layer film having a construction of the system it is attempted.However, intestinal epithelial cells or the equivalent of cells of a single-layer film, an oxygen-free state and reproduce the environment in the intestine, a single-layer film made of the junctions between the cells in the fracture is 1, and 48 within a time cell death because the separation of the single-layer film, the intestinal epithelial cells of the intestinal tract may not be present in the culture (Patent Document 1).
Therefore, so as to mimic the natural intestinal state than the intestinal epithelial cells and bacteria in the intestine and as a co-culture, a porous film of the intestinal epithelial cells were further divided by a micro-channel, divided into upper and lower micro-channel different from the fluid source and continues to be supplied to the culture solution, the micro flow path provided on both sides of the vacuum chamber by a porous film layer of intestinal epithelial cells as stretching, intestinal bacteria culture system of intestinal epithelial cells has been proposed (Patent Document 1).
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • KYOTO UNIVERSITY
  • 発明者(英語)
  • KATAYAMA Takane
  • KAMBE Taiho
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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