- 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
ZUSAMMENFASSUNG
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.
MATERIALIEN
Produktnummer
Marke
Produktbeschreibung
Sigma-Aldrich
Puromycin -dihydrochlorid aus Streptomyces alboniger, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Gelatine aus Schweinehaut, powder, gel strength ~300 g Bloom, Type A, BioReagent, suitable for electrophoresis, suitable for cell culture
Sigma-Aldrich
Mitomycin C aus Streptomyces caespitosus, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
ESGRO® Rekombinantes Maus-LIF-Protein, 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