Mus musculus Raw sequence reads

Stem cell fate is determined by specific niches that provide multiple physical, chemical and biological cues. However, the hierarchy or cascade of impact of these complicated cues remains elusive due to the spatiotemporal complexity of these environments. Here, anisotropic silk protein nanofiber-based hydrogels with cell adhesion capacity were developed to mimic aspects of the physical microenvironment inside the blastocele. Mouse embryonic stem cells (ESCs) maintained a high level of stemness in these biomimetic hydrogels in vitro in the absence of Leukemia Inhibitory Factor (LIF) and embryonic fibroblasts (EFs). The stemness, developmental capacity and germline transmission of ESCs cultured in these hydrogels were superior to those cultured using classical LIF and EFs method. Further, the physical cues of the hydrogels stimulated the secretion of autocrine factors from the ESCs, establishing an improved dynamic niche to maintain stemness. These findings indicate that physical cue is perhaps a key factor in determining cell fate, which controls subsequent chemical and biological cues. Such bioengineered biomaterials appeared to be sufficient to determine the fate of these ESCs. The improved maintenance of ESCs stemness and developmental capacity during long-term cell culture, along with this relatively simple manipulation and low cost option with these protein hydrogels, may help propagate the expansion and utility of ESCs in various applications.