Establishment of the method for non-destructive and non-invasive monitoring of iPS cells developing into EPO producing cells using Raman spectroscopy

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K05556
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 34020:Analytical chemistry-related
• Research Institution: Shimane University
• Principal Investigator: Ishigaki Mika 島根大学, 学術研究院農生命科学系, 助教 (60610871)
• Co-Investigator(Kenkyū-buntansha): 西山 成 香川大学, 医学部, 教授 (10325334)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: ラマン分光法 / iPS細胞 / 分化誘導過程 / エリスロポエチン産生細胞

Outline of Final Research Achievements

We investigated the process of human induced pluripotent stem cells (iPSCs) into erythropoietin (EPO)-producing cells using Raman spectroscopy and imaging. The fixed cells at four stages of cell differentiation (Phase I, II, III, and IV) were analyzed, and the concentration variations of intracellular molecular compositions with the cell differentiation were examined. Three biomarkers for monitoring the cell differentiation were made clear: the bands due to C-S-C stretching vibrational modes, the concentrations of unsaturated fatty acids with low grade of unsaturation, and the concentrations of glycoproteins. The dynamic changes of these molecular compositions during the course of cell differentiation were successfully visualized by Raman imaging. The results showed application possibilities of Raman spectroscopy and imaging to monitor the cell differentiation and discriminate the cell maturation.

Free Research Field

分析化学

Academic Significance and Societal Importance of the Research Achievements

iPS細胞を用いた新しい薬の開発や，再生医療応用への関心は非常に高く，夢の再生医療技術として期待される一方，iPS細胞の安全性の担保や，安定した細胞培養技術の確立など，未だ解決すべき多くの課題がある。本研究は，ラマン分光法を用いたiPS細胞，及び分化誘導細胞の非破壊，非侵襲，ラベルフリー評価法の確立を目的として実施した。iPS細胞がエリスロポエチン産生細胞へ分化する過程において，脂質、糖タンパクの代謝変化を捉えた本結果は世界初の結果であり，学術的意義が非常に高いと言える。また，本手法はiPS細胞の安定的，安全性を担保した細胞培養モニタリング技術への応用が期待され，社会的意義が高いと考えられる。