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Patterning Pluripotent Stem Cells at a Single Cell Level

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Studies of cell-extracellular matrix (ECM) interactions at a single cell level have drawn interest from scientists around the world. Subcellular ECM micropatterning techniques allow researchers to control cell shape, migration, and spindle orientation during mitosis potentially influencing the stem cell fate. Generally these studies have been limited to somatic cells rather than human pluripotent stem cells (hPSCs) which are capable of enormous differentiation potential. hPSCs require a defined ECM for attachment and express characteristic integrins mediating cell-substrate interactions. hPSCs also rely on cell–cell contacts for survival and to maintain self-renewal properties, but these circumstances also significantly limit hPSC observation at a single cell level. In addition, currently available methods for ECM micropatterning generally require a facility with trained personnel and intricate equipment to produce protein micropatterns. To overcome this problem, we have developed a protocol for vitronectin micropatterning using simple UV/ozone modification of polystyrene. Single hPSCs were able to attach and form characteristic stress fibers and focal adhesions similar to somatic cell types which demonstrate hPSC responsiveness to extracellular adhesive cues. Micropatterned hPSCs were able to be cultured for up to 48 hours while maintaining expression of pluripotency-associated transcription factor OCT4. Although further studies are necessary, the results of our investigation will potentially have a large impact on cell regenerative medicine and tissue engineering.
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Keywords: EXTRACELLULAR MATRIX; MICROPATTERNING; PLURIPOTENT STEM CELLS; POLYSTYRENE; STRESS FIBERS; UV/OZONE; VITRONECTIN

Document Type: Research Article

Publication date: 01 August 2013

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  • Journal of Biomaterials and Tissue Engineering (JBT) is an international peer-reviewed journal that covers all aspects of biomaterials, tissue engineering and regenerative medicine. The journal focuses on the broad spectrum of research topics including all types of biomaterials, their properties, bioimplants and medical devices, biofilms, bioimaging, BioMEMS/NEMS, biosensors, fibers, tissue scaffolds, tissue engineering and modeling, artificial organs, tissue interfaces, interactions between biomaterials, blood, cells, tissues, and organs, regenerative medicine and clinical performance.
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