Direct reprogramming to differentiate into hepatocyte-like cells from canine cutaneous fibroblast

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K15050
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science
• Research Institution: Azabu University
• Principal Investigator: Hisasue Masaharu 麻布大学, 獣医学部, 准教授 (80333144)
• Research Collaborator: Suzuki Atsushi ; Neo Sakurako
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: ダイレクトリプログラミング / 骨髄 / 肝臓 / 再生 / ウイルスベクター / 幹細胞 / 皮膚 / イヌ

Outline of Final Research Achievements

In this study, we applied direct reprogramming in differentiating of canine BMSCs by transfecting Foxa1 and Hnf4a into hepatocyte-like cells. The characterization of cBMSC was proved by plastic adherent, differentiation into adipocytes, osteoblasts and chondrocytes, and analysis of cell surface antigens. A virus mixing Foxa 1 and Hnf 4 α was transfected to cBMSCs, and iHep cells was differentiated. Morphology of iHep cells was exhibited circular to equilateral circular shapes. Expression levels of ALB and CDH m-RNA were increased approximately 5,000 and 10,000 times compared to cBMSC, respectively. Protein expression ALB and E-CDH was confirmed by immunohistochemistry, and LDL metabolic ability and Urea production were increased. In the present study, canine BMSCs were successfully induced into functional iHep cells, and they were expected to provide insights into the construction of liver models in drug discovery research and potential therapeutics for liver disease.

Free Research Field

再生医療

Academic Significance and Societal Importance of the Research Achievements

犬では、現在多くの肝疾患が見られるが移植医療をはじめとする根本的な治療法は確立されていない。また、肝組織は長期培養が困難であり、創薬開発のために多数の動物実験を実施せざるをえず、不必要な多くの命が失われている。我々は、ダイレクトリプリグ法という簡便に繊維芽細胞から肝細胞を作成することを目指しさまざまな組織からの肝細胞作成を行ってきた。本研究では皮膚の細胞からの分化を目指したが、最終的には骨髄組織中に存在する細胞群であるcBMSCからの分化誘導が最も効率よく、かつ高品質の肝細胞を作成できた。以上の研究成果は、今後薬剤スクリーニングおよび肝疾患の移植医療発展に大きく貢献するものと考えられた。