- 著者
- Yuki Kondo Sachimi Yoshihashi Kayo Mimori Ruri Ogihara Yoshinori Kanehama Yoshiyuki Maki Shin Enosawa Kouichi Kurose Takahiro Iwao Katsunori Nakamura Tamihide Matsunaga
- 出版者
- 日本薬物動態学会 会長/日本薬物動態学会 DMPK編集委員長
- 雑誌
- Drug Metabolism and Pharmacokinetics (ISSN:13474367)
- 巻号頁・発行日
- pp.DMPK-14-RG-022, (Released:2014-04-29)
- 参考文献数
- 22
- 被引用文献数
- 9
This study aimed to establish culture conditions which are able to give the differentiation of induced pluripotent (iPS) cells to the hepatocytes. To this end, we examined the usefulness of culture medium containing the components involved in the intermediary metabolism in the liver. More specifically, we examined the effect of the 'modified L-15 medium' containing galactose, phenylalanine and ornitine, but deprived of glucose, tyrosine, arginine and pyruvic acid. The medium was altered according to changes in the expression of enzymes that participate in liver-specific pathways. After 25 days of differentiation, the differentiated cells expressed hepatocyte markers and drug-metabolizing enzymes. These expression levels were increased using modified L-15 medium. The survival of human fetal liver cells and the death of human fibroblasts were observed during culture in modified L-15 medium. Most of the cells that differentiated from human iPS cells using modified L-15 medium were stained by anti-human albumin antibody. These results suggest that iPS cells can be converted to high purity-differentiated hepatocytes by cultivating them in modified L-15 medium.
- 著者
- Sandi SUFIANDI Hiromichi OBARA Huai-Che HSU Shin ENOSAWA Hiroshi MIZUNUMA
- 出版者
- The Japan Society of Mechanical Engineers
- 雑誌
- Journal of Biomechanical Science and Engineering (ISSN:18809863)
- 巻号頁・発行日
- pp.17-00421, (Released:2017-12-04)
- 参考文献数
- 34
- 被引用文献数
- 1
Improving the process of cell injection during hepatocyte transplantation requires an understanding of the causal relationships that shear, direct contact cells with a solid surface, and cell deformation have on cell viability loss. A linear shear model was used to model this loss of cell viability during their movement on a solid surface as part of the injection step of hepatocyte transplantation. Rat hepatocytes were studied under linear shear using two parallel plates, with a ”tight” condition that had a 25 μm gap, and a ”loose” gap condition with a > 25 μm gap, to determine the effects of cell deformation, and simulate cell viability loss during injection. Cell morphology and deformation were also observed using time-lapse images. Direct contact with a solid surface is deleterious for cells, and live cells became deformed under shear stress until they lost viability. The cell size could decrease or increase during deformation, and a loss of viability could occur due to a loss of membrane integrity or cell rupture. The space limitations in the tight gap could prevent cell expansion, which delayed the process of cell viability loss. In summary, preventing the direct contact of hepatocytes with a solid surface is recommended to improve the cell injection process during transplantation.
- 著者
- Sandi SUFIANDI Hiromichi OBARA Huai-Che HSU Shin ENOSAWA Naoto MATSUNO Hiroshi MIZUNUMA
- 出版者
- 一般社団法人 日本機械学会
- 雑誌
- Journal of Biomechanical Science and Engineering (ISSN:18809863)
- 巻号頁・発行日
- pp.17-00325, (Released:2017-10-12)
- 参考文献数
- 33
- 被引用文献数
- 5
A sufficient number of functional live hepatocytes delivered to a recipient is necessary for cell therapy. Preventing cell viability loss during the cell injection process is important to improve the clinical outcomes of hepatocyte transplantation. The critical location of cell viability loss is important to identify the causal relationship between the viability loss and cell injection process. In this study, the critical location of cell viability loss was determined experimentally in a rectangular microchannel by microscopic high-speed camera observations. Live hepatocyte distributions were investigated upstream and downstream, and measured on three planes, top, center, and bottom, under horizontally or vertically supplied conditions of the syringe orientation. Sedimented and uniform dispersion conditions of the live hepatocyte distribution at upstream of the microchannel were classified according to observations at horizontal and vertical syringe orientations, respectively. Higher hepatocyte viability loss was found under the sedimented condition. The results suggested that the critical location of hepatocyte viability loss was on the bottom plane of the microchannel. Furthermore, physical causes of the hepatocyte viability loss were found by micro-scale observations of the cell velocity and diameter during the cell injection process. This information may contribute to development of a guideline for the cell injection process to improve hepatocyte transplantation.