Analysis of the behavior of 2D monolayers and 3D spheroid human pancreatic beta cells derived from induced pluripotent stem cells in a microfluidic environment
نویسندگان
چکیده
The limited availability of primary human ?-cells/islets and their quality (due to donor diversity) restrict the development in vitro models for diabetes research. Human induced pluripotent stem cells (hiPSCs) may be a promising cell-source studies, anti-diabetic drug screening personalized therapies. However, achieving levels maturity/functionality that are comparable vivo situation islets rebuilt from iPSCs is still challenging. Here, we compare discuss two strategies culturing pancreatic ?-cells derived hiPSCs microfluidic biochips. First, confirmed protocol conventional Petri 2D monolayer led insulin, PDX1 MAFA positive staining, C-Peptide productive cells, tissue responsive high/low glucose GLP1 stimulation. This its subsequent modifications (including extracellular matrix coating, cell adhesion time, inoculation density, flow rate) was not successful biochip culture. We proposed second strategy using 3D spheroids created honeycomb static cultures. Spheroids experiments carried out over 14 days demonstrated they expressed high ?-cell markers (INS mRNA) higher ?-cell (GCG mRNA glucagon staining), when compared Furthermore, were specifically able secrete insulin response both stimulation exposure. successfully inoculated into biochips maintained 10 perfusion. cultures increased GCG responsiveness Finally, C-peptide secretion spheroids. These results illustrate potential spheroid-based pancreas-on-chip model disease/diabetes modeling screening.
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ژورنال
عنوان ژورنال: Journal of Biotechnology
سال: 2021
ISSN: ['1873-4863', '0168-1656']
DOI: https://doi.org/10.1016/j.jbiotec.2021.02.009