Efficient differentiation of mouse embryonic stem cells into insulin-producing cells.
Embryonic stem (ES) cells are a potential source of a variety of differentiated cells for cell therapy, drug discovery, and toxicology screening. Here, we present an efficacy strategy for the differentiation of mouse ES cells into insulin-producing cells (IPCs) by a two-step differentiation protocol comprising of (i) the formation of definitive endoderm in monolayer culture by activin A, and (ii) this monolayer endoderm being induced to differentiate into IPCs by nicotinamide, insulin, and laminin. Differentiated cells can be obtained within approximately 7 days. The differentiation IPCs combined application of RT-PCR, ELISA, and immunofluorescence to characterize phenotypic and functional properties. In our study, we demonstrated that IPCs produced pancreatic transcription factors, endocrine progenitor marker, definitive endoderm, pancreatic β-cell markers, and Langerhans α and δ cells. The IPCs released insulin in a manner that was dose dependent upon the amount of glucose added. These techniques may be able to be applied to human ES cells, which would have very important ramifications for treating human disease.
Pharmacognosy Lab, Herbal Medicinal Product Technology Division, Industrial Technology Research Institute, Hsinchu 30011, Taiwan. szuhsiuliu@itri.org.tw
Szu-Hsiu
SH
Liu
Lain-Tze
LT
Lee
D000818:Animals; D002454:Cell Differentiation; D046149:Embryo Culture Techniques / Q000379:methods*; D053595:Embryonic Stem Cells / Q000166:cytology*; D004707:Endoderm / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology; D007797:Laminin / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D009536:Niacinamide / Q000378:metabolism; D010641:Phenotype
D007328:Insulin; D007797:Laminin; D053605:Leukemia Inhibitory Factor; D009536:Niacinamide; D005947:Glucose