Two-Stage Structural and Slowing-Down Percolation Transitions in the Densifying Cancer Cell Monolayer.

We experimentally demonstrate the two-stage structural and slowing-down percolating transitions, followed by the confluent transition in the densifying cancer cell monolayers from the dilute state, and investigate their impacts on collective cell dynamics. It is found that cells aggregate into clusters at low cell density. With increasing cell number density, the structural percolation through the formation of a large cell cluster percolating through the space precedes the dynamical percolation transition of forming a percolating cluster of slow cell elements. Both percolating transitions exhibit scale-free scaling behaviors of cluster size distributions and fractal structures, similar to those of the universality class of 2D nonequilibrium systems governed by percolation theory. Dynamically, at low cell density, cell aggregation enhances cooperative motion. The structural percolation leads to slower motion, especially with stronger suppression for the high-frequency modes in the turbulent-like velocity power spectra. The following slowing-down percolation associated with the onset of cell crowding in regions occupied by cells further enhances dynamical slowing-down, and suppresses the increasing trend of dynamical heterogeneity and the steepening of the power spectrum of motion, until their reversions after the confluent transition.

Spontaneous multi-scale void formation and closure in densifying epithelial and fibroblast monolayers from the sub-confluent state.

Using time-lapse phase contrast microscopy, the formation and closure of spontaneously generated voids in the densifying monolayers of isotropic epithelial cells (ECs) and elongated fibroblast cells (FCs) through proliferation from the sub-confluent state are investigated. It is found that, in both types of monolayers after forming a connected network composed of nematic patches with different orientations, numerous multi-scale voids can be spontaneously formed and gradually close with increasing time. The isotropic fluctuations of deformation and crawling of ECs and the anisotropic axial motion/alignment polarizations of FCs are the two keys leading to the following different generic dynamical behaviors. In EC monolayers, voids exhibit irregular boundary fluctuations and easier cell re-orientation of front layer cells (FLCs) surrounding void boundaries. Void closures are mainly through pinching the gap between the opposite fluctuating void boundaries, and the inward crawling of FLCs to reduce void area associated with topological rearrangement to reduce FLC number. In FC monolayers, large voids have piecewise smooth convex boundaries, and cusp-shaped concave boundaries with cells orienting toward the void at cusp tips. The extension of a thin cell bridge from the cusp tip can bisect a large void into smaller voids. For smaller FC voids dominated by convex boundaries, along which cell alignment prohibits inward crawling, the reduction of FLC number through successive outward squeezing of single FLCs by neighboring FLCs sliding along the void boundary plays an important role for topological rearrangement and void closure. Unlike those surrounding artificial wounds in dense EC monolayers, the absence of ring-like purse-strings surrounding EC and FC voids allows topological rearrangements for reducing void perimeter and void area.

Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua.

Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis.