Fabrication of 3D in vitro model of diabetic neuropathy with real-time detection of ROS generation

2020 Fiscal Year Annual Research Report

• Project/Area Number: 20F20361
• Research Institution: The University of Tokyo
• Principal Investigator: 竹内 昌治 東京大学, 大学院情報理工学系研究科, 教授 (90343110)
• Co-Investigator(Kenkyū-buntansha): MYASNIKOVA DINA 東京大学, 情報理工学(系)研究科, 外国人特別研究員
• Project Period (FY): 2020-11-13 – 2023-03-31
• Keywords: skin / vascular structure / perfusion culture

Outline of Annual Research Achievements

From 11.2020 until 04.2021 I mastered the culturing technique for fabrication of skin equivalent (SE) with correct architecture of 4 epidermis layers. I learnt the standard techniques for SE characterization (hematoxylin and eosin staining; measurement of TEER). Now, I am learning the HPLC to perform permeation test, which is an important for characterizing skin barrier function.
I designed a device for fabrication of SE with vascular structure. I optimized the SE culture protocol for the device and learnt perfusion culture technique. Now I am able to fabricate SE with perfusable channels.
Due to the coronavirus the purchasing of iPS cells took longer than I planned, but I am certain that I will be able to start part of the experiment connected with iPS cells from 05.2021.

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

The laboratory has provided all necessary equipment. If some repair or guidance was needed it was provided quickly. Moreover, professor is very enthusiastic at improving research environment, so he helps with finding and choosing a better equipment for required experiment.
I would also like to mention that there are senior researches with a wide background range at the laboratory, who are always helping with setting up the experiment or provide valuable insights and comments to help with the experiment progress.

Strategy for Future Research Activity

Next year goals are to optimizing the perfusion conditions of the SE. And incorporating neuron cells into the SE structure. The last step will require work with iPS cells and may take quite a bit of time, as the culture protocols take at least 1 month until differentiation is complete.
Another important step is to perform a permeation test, as it is vital to assess the SE barrier function and to test the applicability of my model for drug screening.
I will start working with pancreatic islet model and improve it by using iPS cells.
And the last goal of this year would be to fabricated ROS sensing cellulose beads. This will require some background literature work and design of the device for beads fabrication.