Patent ID: 8569061
Filing Date: 2013-10-29
Classification: C12N

Abstract:
1. A method of biasing human embryonic stem cells (hESCs) towards endodermal lineage without substantial differentiation the method comprising: culturing hESCs that were never previously exposed to a supplement of hyaluronidase, wherein the never previously exposed hESCs are cultured in exemplary invention media formulation supplemented with a hyaluronidase concentration that permits biasing towards endodermal lineage, to generate hESCs biased towards endodermal lineage in one of a plurality of increasing concentrations of hyaluronidase, starting from about 0 ug/ml of hyaluronidase and increasing to a concentration of hyaluronidase effective to bias towards endodermal lineage, added to a media formulation that is exemplary invention media formulation, whereby increasing the concentration of hyaluronidase above about 0 ug/ml selectively biases the differential tendency of the hESCs towards endodermal lineage, wherein said exemplary invention media formulation has 1×DMEM/F-12 low osmolality (467.5 mL); 5 mL of 1× non-essential amino acids, 5 mL of 1× Glutamax (L-alanyl-glutamine), 2.5 mL of 0.5× pyruvate, 0.5% human albumin, 20 ug/mL ethanolamine, 5 ug/mL insulin, 10 ug/mL transferrin, 1 ng/mL selenite, 4 ng/mL T3 (thyroid hormone), and 100 uM beta-mercaptoethanol, 494 mL final volume.