title: Laser-assisted photoablation of human pluripotent stem cells from differentiating cultures.
abstract: Due to their pluripotency and their self-renewal capacity, human pluripotent stem cells (hPSC) provide fascinating perspectives for biomedical applications. In the long term, hPSC-derived tissue-specific cells will constitute an important source for cell replacement therapies in non-regenerative organs. These therapeutic approaches, however, will critically depend on the purity of the in vitro differentiated cell populations. In particular, remaining undifferentiated hPSC in a transplant can induce teratoma formation. In order to address this challenge, we have developed a laser-based method for the ablation of hPSC from differentiating cell cultures. Specific antibodies were directed against the hPSC surface markers tumor related antigen (Tra)-1-60 and Tra-1-81. These antibodies, in turn, were targeted with nanogold particles. Subsequent laser exposure resulted in a 98,9 +/- 0,9% elimination of hPSCs within undifferentiated cell cultures. In order to study potential side effects of laser ablation on cells negative for Tra-1-60 and Tra-1-81, hPSC were mixed with GFP-positive hPSC-derived neural precursors (hESCNP) prior to ablation. These studies showed efficient elimination of hPSC while co-treated hESCNP maintained their normal proliferation and differentiation potential. In vivo transplantation of treated and untreated mixed hPSC/hESCNP cultures revealed that laser ablation can dramatically reduce the risk of teratoma formation. Laser-assisted photothermolysis thus represents a novel contact-free method for the efficient elimination of hPSC from in vitro differentiated hPSC-derived somatic cell populations.
authors.affiliation: Institute of Reconstructive Neurobiology, Life & Brain Center, University of Bonn and Hertie Foundation, Sigmund-Freud-Str. 25, 53105, Bonn, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Terstegge
authors.forename: Franziska
authors.initials: F
authors.lastname: Winter
authors.forename: Barbara H
authors.initials: BH
authors.lastname: Rath
authors.forename: Iris
authors.initials: I
authors.lastname: Laufenberg
authors.forename: Claudia
authors.initials: C
authors.lastname: Schwarz
authors.forename: Anke
authors.initials: A
authors.lastname: Leinhaas
authors.forename: Florian
authors.initials: F
authors.lastname: Levold
authors.forename: Andreas
authors.initials: A
authors.lastname: Dolf
authors.forename: Simone
authors.initials: S
authors.lastname: Haupt
authors.forename: Philipp
authors.initials: P
authors.lastname: Koch
authors.forename: Elmar
authors.initials: E
authors.lastname: Endl
authors.forename: Oliver
authors.initials: O
authors.lastname: Brüstle
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D007834:Lasers; D039904:Pluripotent Stem Cells / Q000166:cytology*; D013724:Teratoma / Q000473:pathology