Silencing of p53 and CDKN1A establishes sustainable immortalized megakaryocyte progenitor cells from human iPSCs

• DOI: 10.1016/j.stemcr.2021.11.001
• Publication Date: 2021-12-02T15:30:19Z
• Authors: Masamitsu Sone; Sou Nakamura; Sachiko Umeda; Harumi Ginya; Motohiko Oshima; Maria Alejandra K...; Sudip Kumar Paul; Kanae Hashimoto; Emiri Nakamura; Yasuo Harada; Kyoko Tsujimura; Atsunori Saraya; Tomoyuki Yamaguchi; Naoshi Sugimoto; Akira Sawaguchi; Atsushi Iwama; Koji Eto; Naoya Takayama
• Keywords: p53; Blood Platelets; Cyclin-Dependent Kinase Inhibitor p21; Induced Pluripotent Stem Cells; bcl-X Protein; regenerative medicine; immortalization; Cell Line; megakaryocyte; Proto-Oncogene Proteins c-myc; 03 medical and health sciences; Report; Humans; Gene Silencing; Cell Proliferation; Megakaryocyte Progenitor Cells; platelet; Polycomb Repressive Complex 1; 460; 0303 health sciences; iPSC; cyclin-dependent kinase inhibitor; Clone Cells; Up-Regulation; CDKN1A; HEK293 Cells; Gene Knockdown Techniques; Tumor Suppressor Protein p53

Platelet transfusions are critical for severe thrombocytopenia but depend on blood donors. The shortage of donors and the potential universal HLA-null platelet products have stimulated research ex vivo differentiation human pluripotent stem cells (hPSCs) to platelets. We recently established expandable immortalized megakaryocyte cell lines (imMKCLs) from hPSCs by transducing MYC, BMI1, BCL-XL (MBX). imMKCLs can act as cryopreservable master supply concentrates. However, proliferation rates vary with starting hPSC clone. In this study, we reveal gene expression profiles several MKCL clones that arrest is correlated levels specific cyclin-dependent kinase inhibitors. Silencing CDKN1A p53 overexpression MBX was effective at stably inducing generate functional platelets irrespective Collectively, improvement in generating should contribute industrialization biology.