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Search Results for: Mark S. Anderson returned 6 results
Islet Microphysiological Systems (Acry-Chip & Oxy-Chip)

A Lab-on-a-Chip platform for islets with wide capabilities: long-term culture | controlled & dynamic perifusion | in situ spatio-temporal imaging | serial cell assessment | ease of sample retrieval
FP-3W (3-well fluidific platform)
RRID:SCR_015808
The FP-3W (3-well fluidific platform) is a re-sealable, customizable platform that was optimized for perfusion assays of rodent and human pancreatic islets; it has demonstrated use for GSIS with concomitant live-cell imaging, and optogenetic experiments. Notably, it can be disassembled to retrieve islets for post-processing, and may be suitable as well for long-term culture of islets. Compared with other commercial platforms, the FP-3W has reduced response delay, is more user friendly (simpler), and can be suggested for drug testing because it is PDMS-free. The FP-3W can be integrated with existing instruments such as the PERI4-02 perfusion station and an inverted microscope for imaging.
Microelectrode array for functional testing of pancreatic islets

Microelectrode arrays (MEAs) can be used to better understand the electrophysiology of dissociated islet cells in response to glucose in a real-time, non-invasive method over prolonged culture periods. Human islets dissociated into singular cells can be seeded onto MEA and cultured for up to 7 days. Immunofluorescent imaging revealed that several cellular subtypes of islets remained present after dissociation. At days 1, 3, 5, and 7 of culture, MEA recordings can capture higher electrical activities of islet cells under different glucose concentration conditions, e.g. high glucose vs. low glucose. MEA spiking profiles were similar to the time course of insulin response when glucose concentration is switched from low to high. Extracellular recordings of dissociated islet cells using MEA is an effective approach to rapidly assess islet functionality, and could supplement standard assays such as glucose stimulate insulin response.
Enzymatically crosslinked gelatin-laminin hydrogels for neuromuscular tissue engineering

This uses a water-soluble and non-toxic method to incorporate additional extracellular matrix proteins into gelatin hydrogels, while obviating the use of chemical crosslinkers such as glutaraldehyde. Gelatin hydrogels were fabricated using a range of gelatin concentrations (4%-10%) that corresponded to elastic moduli of approximately 1 kPa-25 kPa, respectively, a substrate stiffness relevant for multiple cell types. Microbial transglutaminase was then used to enzymatically crosslink a layer of laminin on top of gelatin hydrogels, resulting in 2-component gelatin-laminin hydrogels. Human induced pluripotent stem cell derived spinal spheroids readily adhered and rapidly extended axons on GEL-LN hydrogels. Axons displayed a more mature morphology and superior electrophysiological properties on GEL-LN hydrogels compared to the controls. Schwann cells on GEL-LN hydrogels adhered and proliferated normally, displayed a healthy morphology, and maintained the expression of Schwann cell specific markers. Lastly, skeletal muscle cells on GEL-LN hydrogels achieved long-term culture for up to 28 days without delamination, while expressing higher levels of terminal genes including myosin heavy chain, MyoD, MuSK, and M-cadherin suggesting enhanced maturation potential and myotube formation compared to the controls. Future studies will employ the superior culture outcomes of this hybrid substrate for engineering functional neuromuscular junctions and related organ on a chip applications.
Organoid microphysiological system preserves pancreatic islet function within 3D matrix

Three-dimensional (3D) multicellular organoids recapitulate the native complexities of human tissue better than traditional cellular monolayers. As organoids are insufficiently supported using standard static culture, microphysiological systems (MPSs) provide a key enabling technology to maintain organoid physiology in vitro. This polydimethylsiloxane-free MPS enables continuous dynamic culture and serial in situ multiparametric assessments.
Integrated human pseudoislet system and microfluidic platform

The integrated platform combines pseudoislets with a microperifusion system that allows synchronous acquisition of GCaMP6f biosensor signal and hormone secretory profiles; in the original application, this made clear differences in GPCR signaling pathways between human beta and alpha cells.
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