Search bioRxiv⌕ Search

Biology subjects

Mimura, H.

Publications and source records attributed to Mimura, H..

2 recordsLinked to original sources

Cell-based multisensor array for detecting cancer biomarkers in human urine

Detecting volatile urinary biomarkers offers a compelling strategy for noninvasive diagnosis of diseases such as cancer. Biohybrid sensors employing sensor cells that express olfactory receptors--with finely tuned molecular recognition capabilities--are garnering increasing attention. However, their application to physiological samples remains challenging due to variability in sensor cell responsiveness and limited signal intensity. Here, we report a volatile biomarker detection platform based on a multisensor array in which insect olfactory receptor- expressing cells are encapsulated in hydrogel and housed within microwells featuring vertical slits. This structural design enables dense three-dimensional cell immobilization and rapid, uniform exposure to target molecules, enhancing the sensitivity of fluorescence responses and minimizing signal variability. Multiple types of sensor cells on a single array enabled the selective and simultaneous detection of potential cancer-related target molecules--acetophenone, phenol, and 6-methyl-5-hepten-2-one. Furthermore, through hexane extraction combined with gas-phase delivery, we reliably detected acetophenone introduced into human urine at micromolar concentrations. This platform offers a compact, reproducible, and scalable solution for volatile biomarker detection using cell-based sensors.

bioengineering↗

Nuclear assembly in giant unilamellar vesicles encapsulating Xenopus egg extract

The reconstitution of a cell nucleus in a lipid bilayer-enclosed synthetic cell makes great strides in bottom-up synthetic biology. In this study, we propose a method for assembling a nucleus in giant unilamellar vesicles (GUVs). To induce reconstitution of the nucleus, we utilize interphase egg extract of African clawed frogs Xenopus laevis, known as a biochemically controllable cell-free system capable of transforming an added sperm chromatin into a nucleus in vitro. We enhanced GUV formation efficiency by the inverted emulsion method through incorporating prolonged waiting time and adding chloroform into lipid-dispersed oil, facilitating subsequent nuclear assembly reactions in the GUVs.Characterization of nucleus-like structures formed in the GUVs revealed the presence of dense DNA and accumulated GFP-NLS in the structure, indicative of functional nuclear import. Immunostaining further validated the presence of nuclear pore complexes on the surfaces of these nucleus-like structures. Our approach offers a versatile platform for constructing artificial cellular systems that closely mimic eukaryotic cells. TeaserA cell nucleus is reconstituted in lipid bilayer-enclosed confinements using egg extract from African clawed frogs.

synthetic biology↗