Open AccessA Novel Construction of Lentiviral Vectors for Eliminating Tumorigenic Cells from Pluripotent Stem Cells
Author(s) -
Ide Kanako,
Mitsui Kaoru,
Irie Rie,
Matsushita Yohei,
Ijichi Nobuhiro,
Toyodome Soichiro,
Kosai Kenichiro
Publication year - 2018
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.2725
Subject(s) - biology , induced pluripotent stem cell , stem cell , microbiology and biotechnology , viral vector , computational biology , virology , embryonic stem cell , genetics , gene , recombinant dna
The risk of tumor formation poses a challenge for human pluripotent stem cell (hPSC)‐based transplantation therapy. Specific and total elimination of tumorigenic hPSCs by suicide genes (SGs) has not been achieved because no methodology currently exists for testing multiple candidate transgene constructs. Here, we present a novel method for efficient generation of tumorigenic cell‐targeting lentiviral vectors (TC‐LVs) with diverse promoters upstream of a fluorescent protein and SGs. Our two‐plasmid system achieved rapid and simultaneous construction of different TC‐LVs with different promoters. Ganciclovir (GCV) exerted remarkable cytotoxicity in herpes simplex virus thymidine kinase ‐transduced hPSCs, and high specificity for undifferentiated cells was achieved using the survivin promoter (TC‐LV.Surv). Moreover, GCV treatment completely abolished teratoma formation by TC‐LV.Surv‐infected hPSCs transplanted into mice, without harmful effects. Thus, TC‐LV can efficiently identify the best promoter and SG for specific and complete elimination of tumorigenic hPSCs, facilitating the development of safe regenerative medicine. S tem C ells 2018;36:230–239