Efficient generation of liver sinusoidal endothelial-like cells secreting coagulation factor VIII from human induced pluripotent stem cells

Seiji Mitani; Chihiro Hosoda; Yu Onodera; Yoko Takabayashi; Asuka Sakata; Midori Shima; Kohei Tatsumi

doi:10.1016/j.omtm.2024.101355

Efficient generation of liver sinusoidal endothelial-like cells secreting coagulation factor VIII from human induced pluripotent stem cells

Mol Ther Methods Clin Dev. 2024 Oct 18;32(4):101355. doi: 10.1016/j.omtm.2024.101355. eCollection 2024 Dec 12.

Authors

Seiji Mitani¹, Chihiro Hosoda¹, Yu Onodera¹, Yoko Takabayashi¹, Asuka Sakata², Midori Shima², Kohei Tatsumi^{1

2}

Affiliations

¹ Advanced Medical Science of Thrombosis and Hemostasis, Nara Medical University, Kashihara, Nara 634-8521, Japan.
² Medicinal Biology of Thrombosis and Hemostasis, Nara Medical University, Kashihara, Nara 634-8521, Japan.

Abstract

Liver sinusoidal endothelial cells (LSECs) and LSEC progenitor cells (LPCs) derived from human pluripotent stem cells (PSCs) are expected as valuable cell sources for the development of cell therapy for hemophilia A, a congenital deficiency of coagulation factor VIII (FVIII), as LSECs are responsible for FVIII production. However, there is room for improvement in the efficiency of LSEC and LPC differentiation from human PSCs. In this study, we sought to optimize the method of mesoderm differentiation induction, the initial step of LSEC differentiation from human PSCs, to efficiently induce LSEC-like cells capable of secreting FVIII from human induced pluripotent stem cells (iPSCs). Following optimization of the concentration and stimulation period of CHIR99021 (glycogen synthase kinase 3β inhibitor), bone morphogenetic protein 4, fibroblast growth factor 2, and Activin A in the mesoderm induction step, approximately 65% and 54% of cells differentiated into LPCs and LSEC-like cells, respectively. Furthermore, we observed substantial FVIII protein secretion from LSEC-like cells in vitro. In conclusion, we established an efficient method for obtaining LPCs and functional LSEC-like cells from human iPSCs in vitro.

Keywords: coagulation factor VIII; differentiation; hemophilia A; human induced pluripotent stem cell; liver sinusoidal endothelial cell.