ABSTRACT
Cell-based assays have become important tools in the pharmaceutical and biotechnology industries. However, observing and monitoring molecules in cells that mimic the physiological environment is often difficult. Dynamic processes not only increase the accuracy of simulations, but also improve our understanding of the function and regulation of molecules within cells. In this study we used chitosan as a multifunctional biomaterial for selective micropatterning of cells, peptide delivery and covalent bonding with quantum dots (QD) to decrease the cytotoxicity of QD. Our results demonstrate the efficacy of chitosan-QD-peptide-Alexa Fluor 488 in controlling the spread and spatial organization of cells. Cationic chitosan also provided an efficient delivery mechanism to live cells. We used the shift from green to red fluorescence of the chitosan dual color QD peptide to detect biological activity. This methodology has potential applications in high throughput screening of inhibitors and activators of biological mechanisms and pathways and for use in the pharmaceutical industry.
Subject(s)
Chitosan/pharmacology , Fibroblasts/cytology , Fibroblasts/drug effects , Fluorescence Resonance Energy Transfer/methods , Peptides/metabolism , Quantum Dots , 3T3 Cells , Amino Acid Sequence , Animals , Cell Survival/drug effects , Color , Fibroblasts/metabolism , Mice , Molecular Sequence Data , Peptides/chemistry , TransfectionABSTRACT
Stem cells have the surprising potential to develop into many different cell types. Therefore, major research efforts have focused on transplantation of stem cells and/or derived progenitors for restoring depleted diseased cells in degenerative disorders. Understanding the molecular controls, including alternative splicing, that arise during lineage differentiation of stem cells is crucial for developing stem cell therapeutic approaches in regeneration medicine. Alternative splicing to allow a single gene to encode multiple transcripts with different protein coding sequences and RNA regulatory elements increases genomic complexities. Utilizing differences in alternative splicing as a molecular marker may be more sensitive than simply gene expression in various degrees of stem cell differentiation. Moreover, alternative splicing maybe provide a new concept to acquire induced pluripotent stem cells or promote cell-cell transdifferentiation for restorative therapies and basic medicine researches. In this review, we highlight the recent advances of alternative splicing regulation in stem cells and their progenitors. It will hopefully provide much needed knowledge into realizing stem cell biology and related applications.
