Tau EGFP embryonic stem cells: an efficient tool for neuronal lineage selection and transplantation.
Pluripotency and the capacity for continuous self-renewal make embryonic stem (ES) cells an attractive donor source for cell-replacement strategies. A key prerequisite for a therapeutic application of ES cells is the generation of defined somatic cell populations. Here we demonstrate that a targeted insertion of the EGFP gene into the tau locus permits efficient fluorescence-activated cell sorting (FACS)-based lineage selection of ES cell-derived neurons. After in vitro differentiation of heterozygous tau EGFP ES cells into multipotent neural precursors, EGFP is selectively induced in postmitotic neurons of various neurotransmitter phenotypes. By using FACS, ES cell-derived neurons can be enriched to purities of more than 90%. Because neuron-specific EGFP fluorescence is also observed upon transplantation of ES cell-derived neural precursors, the tau EGFP mutant represents a useful tool for the in vivo analysis of grafted ES cell-derived neurons.
Institute of Reconstructive Neurobiology and Department of Neuropathology, University of Bonn Medical Center, Bonn, Germany.
Marius
M
Wernig
Kerry Lee
KL
Tucker
Volker
V
Gornik
Angelika
A
Schneiders
Rachel
R
Buschwald
Otmar D
OD
Wiestler
Yves-Alain
YA
Barde
Oliver
O
Brüstle
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods*; D049452:Green Fluorescent Proteins; D066298:In Vitro Techniques; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008938:Mitosis / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000502:physiology; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D016875:tau Proteins / Q000235:genetics*
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D016875:tau Proteins; D049452:Green Fluorescent Proteins