Autophagic response to cell culture stress in pluripotent stem cells

Autophagy is an important conserved cellular process, both constitutively as a recycling pathway for long lived proteins and as an upregulated stress response. Recent findings suggest a fundamental role for autophagic processes in the maintenance of pluripotent stem cell function. In human embryonic stem cells (hESCS), autophagy was investigated by transfection of LC3-GFP to visualize autophagosomes and with an antibody to LC3B protein. The presence of the primary cilium (PC) in hESCs as the site of recruitment of autophagy-related proteins was also assessed. HESCs (mShef11) in vitro displayed basal autophagy which was upregulated in response to deprivation of culture medium replacement. Significantly higher levels of autophagy were exhibited on spontaneous differentiation of hESCs in vitro. The PC was confirmed to be present in hESCs and therefore may serve to coordinate autophagy function. •Transfection of hESCs with LC3-GFP to visualize autophagosomes and confirmation by immunolocalisation of LC3B protein.•The presence of the primary cilium (PC) in hESCs as the site of recruitment of autophagy-related proteins.•Basal autophagy upregulated in response to deprivation of culture medium replacement.•Significantly higher levels of autophagy exhibited on spontaneous differentiation of hESCs in vitro.•The role of autophagy in pluripotent stem cells summarised.