PT  - JOURNAL ARTICLE
AU  - Mahmoud Khazim
AU  - Lorena Postiglione
AU  - Elisa Pedone
AU  - Dan L. Rocca
AU  - Carine Zahra
AU  - Lucia Marucci
TI  - Towards automated control of embryonic stem cell pluripotency
AID  - 10.1101/685297
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 685297
4099  - http://biorxiv.org/content/early/2019/06/27/685297.short
4100  - http://biorxiv.org/content/early/2019/06/27/685297.full
AB  - Mouse embryonic stem cells (mESCs) have been shown to exist in three distinct pluripotent states (ground, naïve and primed pluripotent states), depending on culture conditions. External feedback control strategies have been, so far, mainly used to automatically regulate gene expression in bacteria and yeast. Here, we exploit a microfluidics/microscopy platform and segmentation and external feedback control algorithms for the automatic regulation of pluripotency phenotypes in mESCs. We show feasibility of automatically controlling, in living mESCs, levels of an endogenous pluripotency gene, Rex1, through a fluorescent reporter, used as control output, and drugs commonly used to modulate pluripotency (i.e. MEK kinase and Gsk3β inhibitors) as control inputs. Our results will ultimately aid in the derivation of superior protocols for pluripotency maintenance and differentiation of mouse and human stem cells.