TY - JOUR

T1 - Changes in Cell Morphology and Actin Organization in Embryonic Stem Cells Cultured under Different Conditions

AU - Barooji, Younes F.

AU - Hvid, Kasper G.

AU - Petitjean, Irene Isturiz

AU - Brickman, Joshua M.

AU - Oddershede, Lene B.

AU - Bendix, Poul M.

PY - 2021

Y1 - 2021

N2 - The cellular cytoskeleton provides the cell with a mechanical rigidity that allows mechanical interaction between cells and the extracellular environment. The actin structure plays a key role in mechanical events such as motility or the establishment of cell polarity. From the earliest stages of development, as represented by the ex vivo expansion of naive embryonic stem cells (ESCs), the critical mechanical role of the actin structure is becoming recognized as a vital cue for correct segregation and lineage control of cells and as a regulatory structure that controls several transcription factors. Naive ESCs have a characteristic morphology, and the ultrastructure that underlies this condition remains to be further investigated. Here, we investigate the 3D actin cytoskeleton of naive mouse ESCs using super-resolution optical reconstruction microscopy (STORM). We investigate the morphological, cytoskeletal, and mechanical changes in cells cultured in 2i or Serum/LIF media reflecting, respectively, a homogeneous preimplantation cell state and a state that is closer to embarking on differentiation. STORM imaging showed that the peripheral actin structure undergoes a dramatic change between the two culturing conditions. We also detected micro-rheological differences in the cell periphery between the cells cultured in these two media correlating well with the observed nano-architecture of the ESCs in the two different culture conditions. These results pave the way for linking physical properties and cytoskeletal architecture to cell morphology during early development.

AB - The cellular cytoskeleton provides the cell with a mechanical rigidity that allows mechanical interaction between cells and the extracellular environment. The actin structure plays a key role in mechanical events such as motility or the establishment of cell polarity. From the earliest stages of development, as represented by the ex vivo expansion of naive embryonic stem cells (ESCs), the critical mechanical role of the actin structure is becoming recognized as a vital cue for correct segregation and lineage control of cells and as a regulatory structure that controls several transcription factors. Naive ESCs have a characteristic morphology, and the ultrastructure that underlies this condition remains to be further investigated. Here, we investigate the 3D actin cytoskeleton of naive mouse ESCs using super-resolution optical reconstruction microscopy (STORM). We investigate the morphological, cytoskeletal, and mechanical changes in cells cultured in 2i or Serum/LIF media reflecting, respectively, a homogeneous preimplantation cell state and a state that is closer to embarking on differentiation. STORM imaging showed that the peripheral actin structure undergoes a dramatic change between the two culturing conditions. We also detected micro-rheological differences in the cell periphery between the cells cultured in these two media correlating well with the observed nano-architecture of the ESCs in the two different culture conditions. These results pave the way for linking physical properties and cytoskeletal architecture to cell morphology during early development.

KW - actin cytoskeleton

KW - super-resolution microscopy (STORM)

KW - embryonic stem cells

KW - primed embryonic stem cells

KW - micro-rheology

KW - cell culturing

KW - optical tweezers

KW - OPTICAL RECONSTRUCTION MICROSCOPY

KW - GROUND-STATE

KW - VISCOELASTICITY

KW - CYTOSKELETON

KW - CORTEX

KW - NAIVE

U2 - 10.3390/cells10112859

DO - 10.3390/cells10112859

M3 - Journal article

C2 - 34831083

VL - 10

JO - Cells

JF - Cells

SN - 2073-4409

IS - 11

M1 - 2859

ER -