Develop a 3D Neurological Disease Model of Human Cortical Glutamatergic Neurons Using Micropillar-Based Scaffolds

Establishing an effective three-dimensional (3D) in vitro culture system to better model human neurological diseases is desirable, since the human brain is a 3D structure. Here, we demonstrated the development of a polydimethylsiloxane (PDMS) pillar-based 3D scaffold that mimicked the 3D microenvironment of the brain. We utilized this scaffold for the growth of human cortical glutamatergic neurons that were differentiated from human pluripotent stem cells. In comparison with the 2D culture, we demonstrated that the developed 3D culture promoted the maturation of human cortical glutamatergic neurons by showing significantly more MAP2 and less Ki67 expression. Based on this 3D culture system, we further developed an in vitro disease-like model of traumatic brain injury (TBI), which showed a robust increase of glutamate-release from the neurons, in response to mechanical impacts, recapitulating the critical pathology of TBI. The increased glutamate-release from our 3D culture model was attenuated by the treatment of neural protective drugs, memantine or nimodipine. The established 3D in vitro human neural culture system and TBI-like model may be used to facilitate mechanistic studies and drug screening for neurotrauma or other neurological diseases

Detection of phosphatidylserine in the plasma membrane of single apoptotic cells using electrochemiluminescence

Phosphatidylserine in the plasma membrane of single apoptotic cells was detected using luminol electrochemiluminescence.</p

Verification of Cellular Cholesterol Efflux through Aqueous Diffusion using Enzyme Modified Microcavity Electrodes

Abstract not Available.</jats:p

Targeted Proteomics Enables Simultaneous Quantification of Folate Receptor Isoforms and Potential Isoform-based Diagnosis in Breast Cancer

AbstractThe distinct roles of protein isoforms in cancer are becoming increasingly evident. FRα and FRβ, two major isoforms of the folate receptor family, generally have different cellular distribution and tissue specificity. However, the presence of FRβ in breast tumors, where FRα is normally expressed, complicates this situation. Prior to applying any FR isoform-based diagnosis and therapeutics, it is essential to monitor the expression profile of FR isoforms in a more accurate manner. An LC-MS/MS-based targeted proteomics assay was developed and validated in this study because of the lack of suitable methodology for the simultaneous and specific measurement of highly homologous isoforms occurring at low concentrations. FRα and FRβ monitoring was achieved by measuring their surrogate isoform-specific peptides. Five human breast cell lines, isolated macrophages and 60 matched pairs of breast tissue samples were subjected to the analysis. The results indicated that FRβ was overexpressed in tumor-associated macrophages (TAMs) but not epithelial cells, in addition to an enhanced level of FRα in breast cancer cells and tissue samples. Moreover, the levels of the FR isoforms were evaluated according to the histology, histopathological features and molecular subtypes of breast cancer. Several positive associations with PR/ER and HER2 status and metastasis were revealed.</jats:p

Electrochemiluminescence Detection and Imaging of Biomolecules at the Single-Cell Level

Electrochemiluminescence (ECL) is an electrochemically induced light produced by the excitation of luminophores in redox reactions. For the past twenty years, ECL analysis has been continuously developed and applied for the sensitive detection of biomolecules at the single-cell level due to its low background interference and the resultant high sensitivity. In recent times, ECL-based microscopy has combined the elements of imaging and has thus emerged as a fast-developed imaging tool to visualize biomolecules in single cells. The surface-confined features of ECL imaging provide detailed information about cell membranes that is not easily obtained using classical fluorescence microscopy. In this review, we summarize the recent works on the detection and imaging of biomolecules at the single-cell level using ECL and discuss the development prospects and challenges in the biological application of this technology in the field of cell analysis