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  • Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells.

Maintenance, Transgene Delivery, and Pluripotency Measurement of Mouse Embryonic Stem Cells.

Methods in molecular biology (Clifton, N.J.) (2015-04-13)
Tetsuya S Tanaka
摘要

This chapter describes standard techniques to (1) maintain mouse embryonic stem cell culture, (2) deliver transgenes into mouse embryonic stem cells mediated by electroporation, nucleofection, lipofection, and retro/lentiviruses, and (3) assess the pluripotency of mouse embryonic stem cells. The last part of this chapter presents induction of random cell differentiation followed by the alkaline phosphatase and embryoid body formation assays, immunofluorescence microscopy, and the teratoma formation assay.

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Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
ESGRO®重组小鼠LIF蛋白, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
Sigma-Aldrich
Anti-Nestin (C-terminal region) antibody produced in rabbit, ~1.5 mg/mL, affinity isolated antibody