Impact of Fibronectin Knockout on Proliferation and Differentiation of Human Infrapatellar Fat Pad-Derived Stem Cells

Fibronectin plays an essential role in tissue development and regeneration. However, the effects of fibronectin knockout (FN1-KO) on stem cells’ proliferation and differentiation remain unknown. In this study, CRISPR/Cas9 generated FN1-KO in human infrapatellar fat pad-derived stem cells (IPFSCs) was evaluated for proliferation ability including cell cycle and surface markers as well as stemness gene expression and for differentiation capacity including chondrogenic and adipogenic differentiation. High passage IPFSCs were also evaluated for proliferation and differentiation capacity after expansion on decellularized ECM (dECM) deposited by FN1-KO cells. Successful FN1-KO in IPFSCs was confirmed by Sanger sequencing and Inference of CRISPR Edits analysis (ICE) as well as immunostaining for fibronectin expression. Compared to the GFP control, FN1-KO cells showed an increase in cell growth, percentage of cells in the S and G2 phases, and CD105 and CD146 expression but a decrease in expression of stemness markers CD73, CD90, SSEA4, and mesenchymal condensation marker CDH2 gene. FN1-KO decreased both chondrogenic and adipogenic differentiation capacity. Interestingly, IPFSCs grown on dECMs deposited by FN1-KO cells exhibited a decrease in cell proliferation along with a decline in CDH2 expression. After induction, IPFSCs plated on dECMs deposited by FN1-KO cells also displayed decreased expression of both chondrogenic and adipogenic capacity. We concluded that FN1-KO increased human IPFSCs’ proliferation capacity; however, this capacity was reversed after expansion on dECM deposited by FN1-KO cells. Significance of fibronectin in chondrogenic and adipogenic differentiation was demonstrated in both FN1-KO IPFSCs and FN(–) matrix microenvironment

Fluorescent Probes for Molecular Imaging of ROS/RNS Species in Living Systems

Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS) are highly reactive species which play crucial roles in many fundamental physiological processes including cellular signalling pathways. Over-production of these reactive species by various stimuli leads to cellular oxidative stress which is linked to various disease conditions. Therefore, the development of novel detection methods for ROS and RNS is of great interest and indispensable for monitoring the dynamic changes of ROS and RNS in cells and for elucidating their mechanisms of trafficking and connections to diseases. We have been recently developing various fluorescent sensors which can selectively detect metal ions, ROS or RNS species in live cells or animals. Our turn-on profluorescent sensors are capable of imaging oxidative stress promoted by metal and H2O2 (i.e. the Fenton Reaction conditions) in living cells (Chem Commun 2010); our highly selective and sensitive iron sensors can image the endogenous exchangeable iron pools and their dynamic changes with subcellular resolution in living neuronal cells (ChemBioChem 2012 and unpublished data), and so do our superoxide sensors (ChemBioChem 2012 and unpublished data). Moreover, we have recently developed nitric oxide (NO) sensors for molecular imaging of stimulated NO production in live cells with subcellular resolution as well as novel near infra red (NIR) sensors for NO imaging in live animals

Observation of Spin Hall Effect in Weyl Semimetal WTe2 at Room Temperature

Discovery of topological Weyl semimetals has revealed the opportunities to realize several extraordinary physical phenomena in condensed matter physics. Specifically, these semimetals with strong spin-orbit coupling, broken inversion symmetry and novel spin texture are predicted to exhibit a large spin Hall effect that can efficiently convert the charge current to a spin current. Here we report the direct experimental observation of a large spin Hall and inverse spin Hall effects in Weyl semimetal WTe2 at room temperature obeying Onsager reciprocity relation. We demonstrate the detection of the pure spin current generated by spin Hall phenomenon in WTe2 by making van der Waals heterostructures with graphene, taking advantage of its long spin coherence length and spin transmission at the heterostructure interface. These experimental findings well supported by ab initio calculations show a large charge-spin conversion efficiency in WTe2; which can pave the way for utilization of spin-orbit induced phenomena in spintronic memory and logic circuit architectures