Progenitor cells derived from gene‐engineered human induced pluripotent stem cells as synthetic cancer cell alternatives for in vitro pharmacology

Abstract

Limitations in genetic stability and recapitulating accurate physiological disease properties challenge the utility of patient-derived (PD) cancer models for reproducible and translational research. A portfolio of isogenic human induced pluripotent stem cells (hiPSCs) with different pan-cancer relevant oncoprotein signatures followed by differentiation into lineage-committed progenitor cells was genetically engineered.Characterization on molecular and biological level validated successful stable genetic alterations in pluripotency state as well as upon differentiation to prove the functionality of our approach. Meanwhile proposing core molecular networks possibly involved

The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H

“…1A, B ). Details on the description of characteristics of our TP53 background mutation cell model, which was applied in this project, can be found in our recently published paper [ 14 ].

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“…Moreover, we used the same IDH1 transfection campaigns in a version of previously described p53- mutation models of iPS11 [ 34 ] using TP53R175H variant (in the future abbreviated as TP53) to establish double factor models. The characterization of our generated, here used TP53 background mutation model was recently described [ 14 ]. Genetic authentication of cells was conducted as previously described [ 14 ].…”

“…The characterization of our generated, here used TP53 background mutation model was recently described [ 14 ]. Genetic authentication of cells was conducted as previously described [ 14 ].…”

“…Establishment of constructs and transduction of glioblastoma (GBM) cell lines U251MG or LN229 stably overexpressing IDH1 or IDH1R132H were performed as previously described [ 38 ]. Genetic authentication of cells was conducted as previously described [ 14 ]. GBM cells were grown in RPMI 1640 medium (Lonza) supplemented with 10% FBS (Thermo Scientific), 1% sodium pyruvate (Thermo Scientific), 185 U/mL penicillin and 185 µg/mL streptomycin (Biochrom) (cell culture medium) at 37 °C in an incubator with humidified air and 5% CO 2 .…”