- 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
PMID25863786
ABSTRACT
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.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Puromycin dihydrochloride from Streptomyces alboniger, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Anti-Nestin (C-terminal region) antibody produced in rabbit, ~1.5 mg/mL, affinity isolated antibody
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.